Protein phosphatase-1 catalytic subunit interactions

ABSTRACT

A method of identifying a compound which modulates the interaction between a PP1c and a regulatory subunit thereof, the method comprising determining whether a compound enhances or disrupts the interaction between (a) a PP1c or a fragment, variant, derivative or fusion thereof or a fusion of a fragment, variant or derivative and (b) a regulatory subunit which is able to bind to PP1c or a PP1c-binding fragment, variant, derivative or fusion of the subunit or a fusion of the fragment, variant or derivative. A method of affecting cellular metabolism or function, the method comprising administering to a cell (a) a compound which modulates the interaction between a PP1c and a regulatory subunit thereof or (b) a compound which mimics the effect of a regulatory subunit of PP1c or (c) a peptide capable of binding a PP1c and which affects the ability of PP1c to bind to a particular target and/or affects the regulation of PP1c activity, or a functional equivalent thereof.

The present invention relates to peptides and protein-proteininteractions and to the use of peptides, peptide analogues and compoundswhich modulate protein-protein interactions in the control of cellularmetabolism and function.

Cellular metabolism or function is controlled by a number of regulatoryagents, which are affected by extracellular factors, for example thephysical condition of the cell or the binding of a messenger molecule toa receptor located on the cell surface. The extracellular factor maythen initiate a cascade of secondary messenger reactions within the cellitself, leading ultimately to changes in some aspects(s) of metabolismor cell function.

It is well recognised by those skilled in the art that phosphorylationor dephosphorylation reactions often play a key role in regulating theactivity of the proteins affected. Dephosphorylation reactions arecatalysed by phosphatase enzymes, the activity of which may themselvesbe controlled by phosphorylation and/or dephosphorylation events. Whilsta substantial amount of knowledge has been accumulated regarding proteinphosphatases as a group, the number and variety of these enzymes is suchthat detailed information concerning the mode of action of a specificphosphatase is not always available. There remains a need to furtherelucidate and characterise particular key enzymes.

The reversible phosphorylation of proteins regulates most aspects ofcell life. About a third of all mammalian proteins are now thought tocontain covalently bound phosphate and since protein kinases andphosphatases probably account for approximately 2-3% of all human geneproducts (Hunter, 1995), many of these enzymes must typicallyphosphorylate/dephosphorylate numerous proteins in vivo. However, it isbecoming increasingly clear that some protein kinases and phosphatasesdo not find their physiological substrates by simple diffusion withincells and that they are frequently directed to particular loci in thevicinity of their substrates by interaction with targeting subunits. Inthis way, the actions of protein kinases and phosphatases withinherently broad specificities are restricted and their propertiestailored to the needs of a particular subcellular location, organelle orprocess (reviewed in Hubbard and Cohen, 1993; Faux and Scott, 1996).

Protein phosphatase-1 (PP1), one of the major protein serine/threoninephosphatases of eukaryotic cells, participates in the control of avariety of cellular functions that include glycogen metabolism, musclecontraction, the exit from mitosis (reviewed in [1,2]) and the splicingof mRNA [3]. However, evidence has been accumulating that differentprocesses are regulated by distinct forms of PP1 in which thephosphatase catalytic subunit (PP1c) is complexed to specific “targetingsubunits”. These proteins not only direct PP1c to particular subcellularlocations, but modify its specificity in unique ways and conferregulation by extracellular agonists (reviewed in [2,3]).

Several targeting subunits have been isolated and characterised,including the G_(M)-subunit that targets PP1c to both the glycogenparticles and sarcoplasmic reticulum of striated muscles [4,5], theG_(L) subunit that targets PP1c to liver glycogen [6,7], the M-complexesresponsible for the association of PP1c with the myofibrils of skeletalmuscle [8,9] and smooth muscle [9-12], the p53 binding protein p53BP2[13] and nuclear proteins such as sds22 [14] and NIPP1 [15,16]. PP1c isalso reported to interact with other mammalian proteins such as theretinoblastoma gene product [17], ribosomal protein L5 [18], a 110 kDanuclear protein that has yet to be identified [15] and two cytosolicproteins, termed inhibitor-1 and inhibitor-2. Inhibitor-1, and itshomologue termed dopamine and cyclic AMP-regulated phosphoprotein(DARPP), become potent PP1 inhibitors after phosphorylation by cyclicAMP-dependent protein kinase (PKA). Inhibitor-1 is thought to inactivatePP1c released from glycogen particles when G_(M) is phosphorylated byPKA [19]. Inhibitor-2 is present as a complex with PP1 in the cytosol,and there is evidence that one of its roles is to act like a molecularchaperone to ensure that the PP1 catalytic centre is folded correctlyprior to its delivery to a specific targeting subunit [20]. It seemslikely that many other PP1-targeting subunits will be identified overthe next few years as a result of the introduction of powerful newtechniques such as microcystin Sepharose affinity chromatography [8] andthe yeast “two hybrid system” [13].

The forms of PP1c isolated so far each contain a single PP1c-bindingsubunit, implying that the interaction of different targeting subunitswith PP1c may be mutually exclusive. This, in turn, suggests that thebinding sites for targeting subunits may overlap, and that theproportion of PP1 directed to any particular location may be determinedby the amounts of each targeting subunit synthesised and their relativeaffinities for PP1. However, the different targeting subunits showsurprisingly little similarity to one another. G_(M) and G_(L) arestructurally related, yet display only 23% amino acid sequence identityover the first 286 residues of G_(M), while G_(L) lacks the C-terminal750 residues of G_(M) [7]. p53BP2 [13] and the M₁₁₀ subunits from smoothmuscle [10,11] and skeletal muscle [8] contain ankyrin repeats, but noother similarities have so far been detected between other PP1 targetingsubunits.

The paradigm for the targeting subunit concept is protein phosphatase-1(PP1), one of the major serine/threonine specific protein phosphatasesof eukaryotic cells (Stralfors el al., 1985). This enzyme is involved incontrolling diverse cellular functions including glycogen metabolism,muscle contraction, the exit from mitosis and the splicing of RNA(Cohen, 1989; Shenolikar, 1994; Wera and Hemmings, 1995). Thesedifferent processes appear to be regulated by distinct PP1 holo-enzymesin which the same catalytic subunit (PP1c) is complexed to differenttargeting or regulatory subunits. The latter class of subunits act toconfer in vivo substrate specificity not only by directing PP1c to thesubcellular loci of its substrates, but also by enhancing or suppressingits activity towards different substrates. In addition, the regulatorysubunits allow the activity of PP1 to be modulated by reversible proteinphosphorylation and second messengers in response to extracellularstimuli.

Many regulatory subunits modulate the activity of PP1 towards itssubstrates. In the instance of the regulatory M₁₁₀ subunit that targetsPP1c to myosin, the region on the M₁₁₀ subunit that enhances thedephosphorylation of myosin by PP1 has now been shown to be distinctfrom the region involved in targetting the PP1-M holoenzyme to myosin.These observations indicate that alterations in the substratespecificity of PP1c are likely to result from conformational changesinduced by interactions with the targetting subunit and not simply as adirect result of targetting PP1c to its substrate. However, in the caseof the glycogen binding subunit G_(M), the dephosphorylation of glycogenphosphorylase and glycogen synthase was enhanced only under conditionswhen both the PP1-G_(M) complex and its substrates were bound toglycogen (Hubbard and Cohen, 1989) suggesting that targetting alone maybe sufficient to enhance specificity.

Whilst the identity of the PP1-binding site(s) on any targeting subunitis unknown, it has now been realised that the control of the substratespecificity and activity of this key regulatory enzyme and itsinteractions are of therapeutic importance. Disruption of PP1-targetingsubunit interactions provide a way of altering selectively the state ofphosphorylation, and hence the activities, of particular PP1 substrates.We have now identified relatively small peptides from the G_(M) andM₁₁₀-subunits that interact with PP1, and which either disrupt or mimicthe distinctive properties of myofibrillar and glycogen-associated formsof PP1. The binding of the G-subunit and the M-subunit of PP1 has alsobeen found to be mutually exclusive.

A first aspect of the invention provides a method of identifying acompound which modulates the interaction between a PP1c and a regulatorysubunit thereof, the method comprising determining whether a compoundenhances or disrupts the interaction between (a) a PP1c or a fragment,variant, derivative or fusion thereof or a fusion of a fragment, variantor derivative and (b) a regulatory subunit which is able to bind to PP1cor a PP1c-binding fragment, variant, derivative or fusion of saidsubunit or a fusion of said fragment, variant or derivative.

Conveniently, the PP1c or a fragment, variant or derivative or fusionthereof or a fusion of a fragment, variant or derivative is one that isproduced using recombinant DNA technology. By “fragment, variant,derivative or fusion of PP1c” we mean any such fragment, variant,derivative or fusion that retains the ability to interact with aregulatory subunit or a suitable PP1c-binding fragment, variant,derivative or fusion of said subunit or a fusion of said fragment,variant or derivative.

By “regulatory subunit” we mean any such regulatory subunit. Furthersubunits are being identified all the time. It is preferred if theregulatory subunit contains the consensus peptide sequence SEQ ID NO:35:Arg/Lys-Val/Ile-Xaa-Phe as described below.

By “PP1c-binding fragment, variant, derivative or fusion of said subunitor a fusion of said fragment, variant or derivative” we include any suchfragments, variants, derivatives and fusions which are able to bind toPP1c. Conveniently, the fragments, variants, derivatives are made usingrecombinant DNA technology or, in the case of peptides and peptidederivatives and analogues they may be made using peptide syntheticmethods.

The enhancement or disruption of the interaction between the said PP1cor a fragment, variant, derivative or fusion thereof or a fusion of afragment, variant or derivative and the said regulatory subunit or afragment, variant, derivative or fusion thereof or a fusion of afragment, variant or derivative can be measured in vitro using methodswell known in the art of biochemistry and including any methods whichcan be used to assess protein-protein, protein-peptide andprotein-ligand interactions.

The said interaction can also be measured within a cell, for exampleusing the yeast two-hybrid system as is well known in the art.

It should be appreciated that before the present invention thedissociation of a PP1c-regulatory subunit has not been achieved using asmall molecule such as a peptide or a peptide analogue or derivative.Thus, it is preferred if the compounds screened in the method of thefirst aspect of the invention are small molecules and in particular thatthey are not intact regulatory subunits of PP1c.

By “small molecule” we include any compounds which have a molecularweight of less than 5000, preferably less than 2000 and more preferablyless than 1000. Conveniently, the compounds screened are compounds whichare able to enter a cell either passively via the cell membrane or viaan active uptake system.

A second aspect of the invention provides a method of identifying acompound which mimics the effect of a regulatory subunit of PP1c, themethod comprising contacting said compound with PP1c and determiningwhether, in the presence of the compound, PP1c adopts the function ofproperties of a PP1c in the presence of a given regulatory subunit.

By “mimics the effect of a regulatory subunit of PP1c” we include themeaning that the compound modifies a property of PP1c in such a way thatPP1c acts, in at least one respect, like PP1c that is interacting with aregulatory subunit.

Examples of the properties of PP1c that may be modified, and examples ofcompounds which modify the properties of PP1c which are thereforeidentifiable in this method are given below.

Preferably, in the methods of the first and second aspects the saidregulatory subunit of PP1c is any one of M₁₁₀, G_(L), G_(M),M-complexes, p53 BP2, sds22, NIPPI, L5, Inhibitor-1, Inhibitor-2, orDARPP.

More preferably, the regulatory subunit of PP1c is any one of M₁₁₀,G_(L), G_(M), M-complexes or p53BP2, and still more preferably theregulatory subunit of PP1c is M₁₁₀ or G_(M).

In relation to the method of the first aspect of the invention thefragment of a regulatory subunit which is able to bind to PP1c is any ofthe peptides C63-T93 of SEQ ID NO:32, G63-N75 of SEQ ID NO:32, E2-R575of SEQ ID NO:34, E2-P243 of SEQ ID NO:34, E2-D118 of SEQ ID NO:34,H10C-P350 of SEQ ID NO:34 and peptide 63-80 of SEQ ID NO:32 G_(M) orfunctional equivalents thereof or peptides comprising said peptidesequences provided that they are not the complete G_(M) regulatorysubunit. Preferably the peptides are not E2-R575 of SEQ ID NO:34 orH100-P350 of SEQ ID NO:34.

As is described in more detail in the Examples, these peptides have beenshown to bind to PP1c and it is convenient, in some circumstances, forthe method to be carried out such that one of these peptide is displacedfrom, or the binding is enhanced to, PP1c. Suitably, the peptide may belabelled in a detectable manner to facilitate the detection of theinteraction with PP1c. Conveniently, the peptide is labelledradioactively or fluorescently using methods well known in the art.

Also in relation to the method of the first aspect of the invention thefragment of a regulatory subunit which is able to bind to PP1c is any ofthe peptides M1-E309 of SEQ ID NO:33, M1-F38 of SEQ ID NO:33, M1-A150 ofSEQ ID NO:33 or L24-Y496 of SEQ ID NO:33 of M₁₁₀ or functionalequivalents thereof or peptides comprising said peptide sequencesprovided that they are not the complete M₁₁₀ regulatory subunit.

As is shown in more detail in the Examples these peptides have beenshown to bind to PP1c.

Also in relation to the first aspect of the invention the PP1c-bindingfragment, variant or derivative of said regulatory subunit or a fusionof said fragment, variant or derivative comprises the consensus peptidesequence SEQ ID NO:35: Arg/Lys-Val/Ile-Xaa-Phe wherein Xaa is any aminoacid.

We have found that, surprisingly, many regulatory subunits that bind toPP1c contain the consensus peptide sequence Arg/Lys-Val/Ile-Xaa-Phewherein Xaa is any amino acid, preferably a naturally occurring aminoacid. Typically, the PP1c-binding fragment, variant or derivative ofsaid regulatory subunit or a fusion of said fragment, variant orderivative is a peptide (typically 8-400 amino acid residues, preferably8-200, more preferably 8-10 and still more preferably 8-20 amino acidresidues in length which comprises the given consensus peptidesequence).

It is preferred if the PP1c-binding fragment, variant or derivativecomprises, in addition to the said consensus peptide sequence, at leastone basic residue in the four residues N-terminal of the consensuspeptide sequence. Preferably, there are at least two basic residues inthis position, more preferably at least three such residues.

It is also preferred wherein in the consensus peptide sequence Xaa isnot Asp or Glu because the negative charge is believed to interfere withbinding to PP1c. Similarly, it is preferred if Xaa is not a largehydrophobic residue such as Phe, Tyr, Trp, lie or Leu.

It is particularly preferred if the PP1c-binding fragment is a fragmentof a regulatory subunit comprising the said consensus peptide sequenceand therefore the peptide sequences which flank the consensus peptidesequence are the same as in the native regulatory subunit.

Preferably the PP1c-binding fragment is a fragment of any of the M₁₁₀,G_(L), G_(M), M-complexes, p53BP2, sds22, NIPPI, L5, Inhibitor-1,Inhibitor-2 or DARPP regulatory subunits comprising said consensussequence.

Although the methods of the first and second aspects of the invention donot rely on any particular mechanism whereby the modulation or mimickingoccurs, it is preferred if the compound binds to a PP1c. Alternatively,but still preferably, the compound binds to a regulatory subunit ofPP1c.

A further aspect of the invention provides a compound identifiable bythe method of the first or second aspects of the invention.

A further aspect of the invention provides a compound which modulatesthe interaction between a PP1c and a regulatory subunit thereof saidcompound comprising any of the peptides G63-T93 of SEQ ID NO:32, G63-N75of SEQ ID NO:32, E2-R575 of SEQ ID NO:34, E2-P243 of SEQ ID NO:34,E2-D118 of SEQ ID NO:34, H100-P350 of SEQ Or NO:34 and peptide 63-80 ofSEQ ID NO:32 G_(M) or functional equivalents thereof or said compoundcomprising any of the peptides M1-E309 of SEQ ID NO:33, M1-F38 of SEQ IDNO:33, M1-A150 of SEQ ID NO:33 or L24-Y496 of SEQ ID NO:33 of M₁₁₀ orfunctional equivalents thereof or said compound comprising the consensuspeptide sequence SEQ ID NO:35: Arg/Lys-Val/Ile-Xaa-Phe wherein Xaa isany naturally occurring amino acid or functional equivalents thereof,provided that the said compound is no a complete regulatory subunit ofPP1c. Preferably the peptides are not E2-R575 of SEQ ID NO:34 orH100-P350 of SEQ ID NO:34.

By “functional equivalent” we include the meaning that the compound,although having a different structure to the said peptides, modulatesthe interaction between a PP1c and a regulatory subunit thereof insubstantially the same way. For example, a functional equivalent may bea peptide in which conservative substitutions have been made. By“conservative substitution” is intended combinations such as Gly, Ala;Val, Ile, Leu; Asp, Glu; Asn, Gln; Ser, Thr; Lys, Arg; and Phe, Tyr. Afunctional equivalent may also be a peptide with the given sequencewhich has been adapted to be more likely to enter a cell. For example,fatty acids or other hydrophobic moieties may be attached to thepeptide.

By the term “peptide” we mean derivatives of peptides which areresistant to proteolysis, for example those in which the N or C terminiare blocked, or both are blocked, and it includes molecules in which oneor more of the peptide linkages are modified so that the moleculeretains substantially the same molecular configuration in the linkagebut the linkage is more resistant to hydrolysis than a peptide linkage.

It is particularly preferred if the compound consists of the peptidesG63-T93 of SEQ ID NO:32, G63-N75 of SEQ ID NO:32, E2-R575 of SEQ IDNO:34, E2-P243 of SEQ ID NO:34, E2-D118 of SEQ ID NO:34, H100-P350 ofSEQ ID NO:34 or peptide 63 to 80 of SEQ ID NO:32 G_(M) or functionalequivalents thereof or if the compound consists of the peptides M1-E309of SEQ ID NO:33, M1-F38 of SEQ ID NO:33, M1-A150 of SEQ ID NO:33 orL24-Y496 of SEQ ID NO:33 of M₁₁₀ or functional equivalents thereof.Preferably, the peptide is not E2-R575 of SEQ ID NO:34 or H100-P350 ofSEQ ID NO:34.

A still further aspect of the invention provides a method of identifyinga compound which modulates the interaction between a PP1c and aregulatory subunit thereof, or binds PP1c or mimics the effect of aregulatory subunit, the method comprising selecting a compound which iscapable of adopting the same or substantially the same conformation as apeptide bound to the regulatory subunit-binding site of PP1c or the sameor substantially the same conformation as the portion of PP1c whichbinds to said peptide. Suitably, the peptide comprises the consensuspeptide sequence Arg/Lys-Val/Ile-Xaa-Phe wherein Xaa is any amino acid,preferably a naturally occurring amino acid. Conveniently, the saidpeptide consists of residues 63 to 75 of G_(M).

It is particularly preferred if the conformation of the said peptide andthe conformation of the said portion of PP1c is as defined by referenceto the atomic coordinates given in Table A (see also Example 2). Example2 provides further details of the peptide—PP1c interactions.

Table A provides the atomic coordinates for the given PP1c-peptidecrystal structure.

A further aspect of the invention provides a compound identifiable bythe aforementioned method of the invention.

It will be appreciated that the aforementioned compounds and peptideswill be useful in medicine and, accordingly, the invention includespharmaceutical compositions of the said compounds in combination with apharmaceutically acceptable carrier.

The formulations may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacy.Such methods include the step of bringing into association the activeingredient (compound of the invention) with the carrier whichconstitutes one or more accessory ingredients. In general theformulations are prepared by uniformly and intimately bringing intoassociation the active ingredient with liquid carriers or finely dividedsolid carriers or both, and then, if necessary, shaping the product.

Formulations in accordance with the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets, each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. The active ingredient mayalso be presented as a bolus, electuary or paste.

A tablet may be made by compression or moulding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active ingredient in afree-flowing form such as a powder or granules, optionally mixed with abinder (eg povidone, gelatin, hydroxypropylmethyl cellulose), lubricant,inert diluent, preservative, disintegrant (eg sodium starch glycolate,cross-linked povidone, cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Moulded tablets may be made bymoulding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. The tablets may optionally becoated or scored and may be formulated so as to provide slow orcontrolled release of the active ingredient therein using, for example,hydroxypropylmethylcellulose in varying proportions to provide desiredrelease profile.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavoured basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouth-washes comprising the active ingredient in asuitable liquid carrier.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents. The formulations may be presented in unit-dose or multi-dosecontainers, for example sealed ampoules and vials, and may be stored ina freeze-dried (lyophilised) condition requiring only the addition ofthe sterile liquid carrier, for example water for injections,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Preferred unit dosage formulations are those containing a daily dose orunit, daily sub-dose or an appropriate fraction thereof, of an activeingredient.

It should be understood that in addition to the ingredients particularlymentioned above the formulations of this invention may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavouring agents.

A further aspect of the invention provides a method of affectingcellular metabolism or function, the method comprising administering toa cell (a) a compound which modulates the interaction between a PP1c anda regulatory subunit thereof or (b) a compound which mimics the effectof a regulatory subunit of PP1c or (c) a peptide capable of binding aPP1c and which affects the ability of PP1c to bind to a particulartarget and/or affects the regulation of PP1c activity, or a functionalequivalent thereof.

It will be appreciated that the said compounds are disclosed above withrespect to specific compounds or with respect to methods of obtainingsuch compounds.

In particular, it is preferred if the compound administered to the cellis any one or more of the peptides G63-T93 of SEQ ID NO:32, G63-N75 ofSEQ ID NO:32, E2-R575 of SEQ ID NO:34, E2-P243 of SEQ ID NO:34, E2-D118of SEQ ID NO:34, H100-P350 of SEQ ID NO:34 and peptide 63-80 of SEQ IDNO:32 G_(M) or functional equivalents thereof or peptides comprisingsaid peptide sequences or any one or more of the peptides M1-E309 of SEQID NO:33, M1-F38 of SEQ ID NO:33, M1-A150 of SEQ ID NO:33 or L24-Y496 ofSEQ ID NO:33 of M₁₁₀ or functional equivalents thereof or peptidescomprising said peptide sequences. Preferably, the peptide is notE2-R575 of SEQ ID NO:34 or H100-P35C of SEQ ID NO:34.

In this embodiment it will be appreciated that functional equivalentsinclude those compounds defined above as being functional equivalents,in particular, derivatives of peptides which are more readily able toenter a cell.

The compound may be administered to the cell in any suitable way, inparticular in such a way that the compound will enter the cell in asuitable form to have its desired effect. Method of facilitating theentry of a compound into the cell are known in the art, for example, inrelation to peptides the importins and penetrations may be used, or thepeptides may be micro-injected or they may enter the cell in a suitablevehicle such as in a liposome.

Preferably, the cell is a cell in a mammalian body.

The aforementioned compounds of the invention or a formulation thereofmay be administered by any conventional method including oral andparenteral (eg subcutaneous or intramuscular) injection. The treatmentmay consist of a single dose or a plurality of doses over a period oftime.

Whilst it is possible for a compound of the invention to be administeredalone, it is preferable to present it as a pharmaceutical formulation,together with one or more acceptable carriers. The carrier(s) must be“acceptable” in the sense of being compatible with the compound of theinvention and not deleterious to the recipients thereof. Typically, thecarriers will be water or saline which will be sterile and pyrogen free.

A further aspect of the invention provides a method of treating apatient in need of modulation of PP1c activity or function the methodcomprising administering to the patient an effective amount of acompound which modulates the interaction between a PP1c and a regulatorysubunit thereof or (b) a compound which mimics the effect of aregulatory subunit of PP1c or (c) a peptide capable of binding a PP1cand which affects the ability of PP1c to bind to a particular targetand/or affects the regulation of PP1c activity, or a functionalequivalent thereof.

As will be apparent from what is described herein, protein phosphatase-1(PP1) is one of the principal serine/threonine-specific proteinphosphatases in human cells where it plays key roles in regulating avariety of physiological roles, including the metabolism of glycogen,the splicing of mRNA, the exit from mitosis and the contraction ofsmooth muscle. The different functions of PP1 are carried out bydistinct species of this enzyme in which the same catalytic unit iscomplexed to different “targeting” subunits. The latter class ofproteins direct PP1 to specific subcellular loci, tailor its propertiesto the needs of a particular locus and confer the ability to beregulated by extracellular signals (hormones, growth factors,neurotransmitters). Compounds as herein described that disrupt specificPP1-“targeting” subunits interactions or mimic the effect of a targetingsubunit are likely to have a number of therapeutic uses as outlinedbelow.

PP1 interacts with the M110-subunit which targets it to myosin in smoothmuscle and enhances the rate at which PP1 dephosphorylates myosin. Thedephosphorylation of myosin underlies the relaxation of smooth muscle.Thus compounds such as those disclosed herein which disrupt theinteraction of PP1 with M110 in arterial muscle are expected to increasethe phosphorylation of arterial myosin and elevate blood pressure.

The interaction of PP1 with M110 enhances the rate at which PP1dephosphorylates myosin, but suppresses the rate at which itdephosphorylates glycogen phosphorylase. The disruption of the PP1-M110interaction is therefore measured in a screen by looking for compoundswhich enhance the dephosphorylation of phosphorylase and/or suppress thedephosphorylation of the myosin P-light chain (see the Examples).

Compounds, such as those disclosed herein, that mimic the effect of theM110 subunit in stimulating myosin dephosphorylation are expected to beuseful in lowering blood pressure. Such compounds are identified bytheir ability to stimulate the dephosphorylation of the myosin P-lightchain by the catalytic subunit of PP1. An example of such an assay,which shows that the N-terminal 38 residues of the M110 subunitstimulate the dephosphorylation of the myosin P-light chain by PP1, isshown in the Examples.

The interaction of PP1 with G_(L) targets the phosphatase to liverglycogen. This interaction enhances the dephosphorylation glycogensynthase which stimulates the conversion of glucose to glycogen. Acompounds, such as those disclosed herein, disrupts the interactionbetween PP1 and G_(L) is expected to be useful in treating hypoglycemia.The interaction of G_(L) with PP1 strongly suppresses the rate at whichPP1 dephosphorylates glycogen phosphorylase. A compound, such as thosedisclosed herein, which disrupts the interaction of PP1 with G_(L) canbe screened for very simply by its ability to increase the phosphorylasephosphatase activity of PP1 G_(L). This can be carried out, for example,using rat liver glycogen pellet as described in the Examples. There isno need to use the purified enzyme.

PP1 interacts with p53 BP2 (Helps et al, 1995) a protein which is knownto interact with the tumour suppressor p53. The phosphorylation of p53is known to enhance its ability to bind to DNA and hence its ability tofunction as a tumour suppressor. p53BP2 may be a protein which targetsPP1 to p53 stimulating the dephosphorylation and inactivation of p53. Acompound, such as those disclosed herein, which disrupts the interactionof PP1 with p53BP2 may enhance the phosphorylation of p53 and itsability to function as a tumour suppressor. Since p53BP2 suppresses thedephosphorylation of glycogen phosphorylase (Helps et al, 1995),compounds that disrupt the p53BP2-PP1 complex can be screened bymeasuring the increase in rate of dephosphorylation of glycogenphosphorylase.

The present invention provides peptides able to bind to the catalyticsub-unit of protein phosphatase-1 (hereinafter referred to as PP1c).Generally the peptides affect the ability of PP1c to bind to particulartarget(s) and/or the regulation of PP1c activity.

Peptides can be designed based on the sequences of regulatory subunits,especially in relation to the peptide consensus sequence found thereinand its flanking sequences. Peptides can be synthesised by methods wellknown in the art. For example, peptides may be synthesised by theFmoc-polyamide mode of solid-phase peptide synthesis as disclosed by Luet al (1981) J. Org. Chem. 46, 3433 and references therein. TemporaryN-amino group protection is afforded by the 9-fluorenylmethyloxycarbonyl(Fmoc) group. Repetitive cleavage of this highly base-labile protectinggroup is effected using 20% piperidine in N,N-dimethylformamide.Side-chain functionalities may be protected as their butyl ethers (inthe case of serine threonine and tyrosine), butyl esters (in the case ofglutamic acid and aspanic acid), butyloxycarbonyl derivative (in thecase of lysine and histidine), trityl derivative (in the case ofcysteine) and 4-methoxy-2,3,6-trimethylbenzenesulphonyl derivative (inthe case of arginine). Where glutamine or asparagine are C-terminalresidues, use is made of the 4,4′-dimethoxybenzhydryl group forprotection of the side chain amido functionalities. The solid-phasesupport is based on a polydimethyl-acrylamide polymer constituted fromthe three monomers dimethylacrylamide (backbone-monomer),bisacryloylethylene diamine (cross linker) and acryloylsarcosine methylester (functionalising agent). The peptide-to-resin cleavable linkedagent used is the acid-labile 4-hydroxymethyl-phenoxyacetic acidderivative. All amino acid derivatives are added as their preformedsymmetrical anhydride derivatives with the exception of asparagine andglutamine, which are added using a reversedN,N-dicyclohexyl-carbodiimide/1-hydroxybenzotriazole mediated couplingprocedure. All coupling and deprotection reactions are monitored usingninhydrin, trinitrobenzene sulphonic acid or isotin test procedures.Upon completion of synthesis, peptides are cleaved from the resinsupport with concomitant removal of side-chain protecting groups bytreatment with 95% trifluoroacetic acid containing a 50% scavenger mix.Scavengers commonly used are ethanedithiol, phenol, anisole and water,the exact choice depending on the constituent amino acids of the peptidebeing synthesised. Trifluoroacetic acid is removed by evaporation invacuo, with subsequent trituration with diethyl ether affording thecrude peptide. Any scavengers present are removed by a simple extractionprocedure which on lyophilisation of the aqueous phase affords the crudepeptide free of scavengers. Reagents for peptide synthesis are generallyavailable from Calbiochem-Novabiocbem (UK) Ltd, Nottingham NG7 2QJ, UK.Purification may be effected by any one, or a combination of, techniquessuch as size exclusion chromatography, ion-exchange chromatography and(principally) reverse-phase high performance liquid chromatography.Analysis of peptides may be carried out using thin layer chromatography,reverse-phase high performance liquid chromatography, amino-acidanalysis after acid hydrolysis and by fast atom bombardment (FAB) massspectrometric analysis.

The peptides may be derived from the targeting subunit(s) of PP1c, inparticular from the subunits G_(L), G_(M), M₁₁₀ and/or M₂₁. Additionallythe peptides may be derived from other subunits such as differentM-complexes, p53BP2, sds22, NIPP1, L5, Inhibitor-1, Inhibitor-2, DARPPor the like. Functional equivalents or portions of these peptides mayalso be used.

In a further aspect the present invention provides the use of peptidesderived from targeting subunit(s) of PP1c, functional equivalents orportions thereof to affect cellular metabolism or function.

In a further aspect the present invention provides a method of treatmentof the human or non-human (preferably mammalian) animal body, saidmethod comprising altering the levels of peptides derived from targetingsubunit(s) of PP1c, functional equivalents or portions thereof to anextent that cellular metabolism or function is affected.

Aspects of cellular metabolism that may be affected include (but are notlimited to) glycogen metabolism, muscle metabolism, physiology andfunction.

Generally the levels of peptides or their activity will be enhanced incells and this control may be achieved by causing higher levels ofexpression of nucleotides sequences encoding for such peptides(optionally linked to molecules which allow them to cross a cellmembrane) or through the administration of such peptides or precursorsthereof. Alternatively, in some circumstances, it may be more desirableto depress the levels of certain peptides or at least to depress thelevel of peptides in active form.

Preferred peptides according to the present invention are derivatives ofG_(M), especially G63-T93 of SEQ ID NO:32, G63-N75 of SEQ ID NO:32,E2-R575 of SEQ ID NO:34, E2-P243 of SEQ ID NO:34, E2-D118 of SEQ IDNO:34, H100-P350 of SEQ ID NO:34 and peptide 63 to 80 of SEQ ID NO:32,and derivatives of M₁₁₀, especially M1-E309 of SEQ ID NO:33, M1-F38 ofSEQ ID NO:33, M1-Al5C of SEQ ID NO:32, and L24-Y496 of SEQ ID NO:33.Preferably, the peptide is not E2-R575 of SEQ ID NO:34 or H100-P35C ofSEQ ID NO:34.

Particularly preferred peptides are those derived from amino acid nos.63 to 93 of SEQ ID NO:32, (including 63-80 and 63-75) of G_(M);or fromamino acids 1 to 309 of SEQ ID NO:33 (including from 1-150 and 1-38) ofM₁₁₀.

The sequence of G_(M) is given in Chen et al (1994) Diabetes 43,1234-1241.

In yet further aspect the present invention provides chimeric proteinscontaining portions of other proteins or peptides or containingadditional amino acids.

Additionally the present invention provides nucleotide sequences(optionally in the form of plasmids) encoding the peptides or chimericproteins of interest. DNA which encodes the polypeptides or peptides ofthe invention or chimeric proteins can be made based on a knowledge ofthe peptide sequences disclosed herein. The DNA is then expressed in asuitable host to produce a polypeptide comprising the compound of theinvention. Thus, the DNA encoding the polypeptide constituting thecompound of the invention may be used in accordance with knowntechniques, appropriately modified in view of the teachings containedherein, to construct an expression vector, which is then used totransform an appropriate host cell for the expression and production ofthe polypeptide of the invention. Such techniques include thosedisclosed in U.S. Pat. No. 4,440,859 issued 3 Apr. 1984 to Rutter et at,U.S. Pat. No. 4,530,901 issued 23 Jul. 1985 to Weissman, U.S. Pat. No.4,582,800 issued 15 Apr. 1986 to Crowl, U.S. Pat. No. 4,677,063 issued30 Jun. 1987 to Mark et at, U.S. Pat. No. 4,678,751 issued 7 Jul. 1987to Goeddel, U.S. Pat. No. 4,704,362 issued 3 Nov. 1987 to Itakura et at,U.S. Pat. No. 4,710,463 issued 1 Dec. 1987 to Murray, U.S. Pat. No.4,757,006 issued 12 Jul. 1988 to Toole, Jr. et at, U.S. Pat. No.4,766,075 issued 23 Aug. 1988 to Goeddel et at and U.S. Pat. No.4,810,648 issued 7 Mar. 1989 to Stalker, all of which are incorporatedherein by reference.

The DNA encoding the polypeptide constituting the compound of theinvention may be joined to a wide variety of other DNA sequences forintroduction into an appropriate host. The companion DNA will dependupon the nature of the host, the manner of the introduction of the DNAinto the host, and whether episomal maintenance or integration isdesired.

Generally, the DNA is inserted into an expression vector, such as aplasmid, in proper orientation and correct reading frame for expression.If necessary, the DNA may be linked to the appropriate transcriptionaland translational regulatory control nucleotide sequences recognised bythe desired host, although such controls are generally available in theexpression vector. The vector is then introduced into the host throughstandard techniques. Generally, not all of the hosts will be transformedby the vector. Therefore, it will be necessary to select for transformedhost cells. One selection technique involves incorporating into theexpression vector a DNA sequence, with any necessary control elements,that codes for a selectable trait in the transformed cell, such asantibiotic resistance. Alternatively, the gene for such selectable traitcan be on another vector, which is used to co-transform the desired hostcell.

Host cells that have been transformed by the recombinant DNA of theinvention are then cultured for a sufficient time and under appropriateconditions known to those skilled in the art in view of the teachingsdisclosed herein to permit the expression of the polypeptide, which canthen be recovered.

Many expression systems are known, including bacteria (for example E.coli and Bacillus subtilis), yeasts (for example Saccharomycescerevisiae), filamentous fungi (for example Aspergillus), plant cells,animal cells and insect cells.

The vectors include a prokaryotic replicon, such as the ColE1 ori, forpropagation in a prokaryote, even if the vector is to be used forexpression in other, non-prokaryotic, cell types. The vectors can alsoinclude an appropriate promoter such as a prokaryolic promoter capableof directing the expression (transcription and translation) of the genesin a bacterial host cell, such as E. coli, transformed therewith.

A promoter is an expression control element formed by a DNA sequencethat permits binding of RNA polymerase and transcription to occur.Promoter sequences compatible with exemplary bacterial hosts aretypically provided in plasmid vectors containing convenient restrictionsites for insertion of a DNA segment of the present invention.

Typical prokaryotic vector plasmids are pUC18, pUC19, pBR322 and pBR329available from Biorad Laboratories, (Richmond, Calif., USA) and pTrc99Aand pKK223-3 available from Pharmacia, Piscataway, N.J., USA.

A typical manimalian cell vector plasmid is pSVL available fromPharmacia, Piscataway, N.J., USA. This vector uses the SV40 latepromoter to drive expression of cloned genes, the highest level ofexpression being found in T antigen-producing cells, such as COS-1cells.

An example of an inducible mammalian expression vector is pMSG, alsoavailable from Pharmacia. This vector uses the glucocorticoid-induciblepromoter of the mouse mammary tumour virus long terminal repeat to driveexpression of the cloned gene.

Useful yeast plasmid vectors are pRS403-406 and pRS413-416 and aregenerally available from Stratagene Cloning Systems, La Jolla, Calif.92037, USA. Plasmids pRS403, pRS404, pRS405 and pRS406 are YeastIntegrating plasmids (YIps) and incorporate the yeast selectable markersHIS3, TRP1, LEU2 and URA3. Plasmids pRS413-416 are Yeast Centromereplasmids (YCps).

A variety of methods have been developed to operably link DNA to vectorsvia complementary cohesive termini. For instance, complementaryhomopolymer tracts can be added to the DNA segment to be inserted to thevector DNA. The vector and DNA segment are then joined by hydrogenbonding between the complementary homopolymeric tails to formrecombinant DNA molecules.

Synthetic linkers containing one or more restriction sites provide analternative method of joining the DNA segment to vectors. The DNAsegment, generated by endonuclease restriction digestion as describedearlier, is treated with bacteriophage T4 DNA polymerase or E. coli DNApolymerase I, enzymes that remove protruding, 3′-single-stranded terminiwith their 3′-5′-exonucleolytic activities, and fill in recessed 3′-endswith their polymerizing activities.

The combination of these activities therefore generates blunt-ended DNAsegments. The blunt-ended segments are then incubated with a large molarexcess of linker molecules in the presence of an enzyme that is able tocatalyze the ligation of blunt-ended DNA molecules, such asbacteriophage T4 DNA ligase. Thus, the products of the reaction are DNAsegments carrying polymeric linker sequences, at their ends. These DNAsegments are then cleaved with the appropriate restriction enzyme andligated to an expression vector that has been cleaved with an enzymethat produces termini compatible with those of the DNA segment.

Synthetic linkers containing a variety of restriction endonuclease sitesare commercially available from a number of sources includingInternational Biotechnologies Inc, New Haven, Conn., USA.

A desirable way to modify the DNA encoding the polypeptide of theinvention is to use the polymerase chain reaction as disclosed by Saikiet al (1988) Science 239, 487-491.

In this method the DNA to be enzymatically amplified is flanked by twospecific oligonucleotide primers which themselves become incorporatedinto the amplified DNA. The said specific primers may containrestriction endonuclease recognition sites which can be used for cloninginto expression vectors using methods known in the art. In relation tothe above section on DNA expression the term “polypeptide” includespeptides and chimeric proteins.

Further the present invention provides host cells transformed withsuitable expression vectors and able to express the peptides. The hostcells may be prokaryotic (e.g. E. coli) or eukaryotic (e.g. yeast,mammalian cell cultures).

Bacterial cells are preferred prokaryotic host cells and typically are astrain of E. coli such as, for example, the E. coli strains DH5available from Bethesda Research Laboratories Inc., Bethesda, Md., USA,and RR1 available from the American Type Culture Collection (ATCC) ofRockville, Md., USA (No ATCC 31343). Preferred eukaryotic host cellsinclude yeast and mammalian cells, preferably vertebrate cells such asthose from a mouse, rat, monkey or human fibroblastic cell line. Yeasthost cells include YPH499, YPH500 and YPH501 which are generallyavailable from Stratagene Cloning Systems, La Jolla, Calif. 92037, USA.Preferred mammalian host cells include Chinese hamster ovary (CHO) cellsavailable from the ATCC as CCL61, NIH Swiss mouse embryo cells NIH/3T3available from the ATCC as CRL 1658, and monkey kidney-derived COS-1cells available from the ATCC as CRL 1650.

Transformation of appropriate cell hosts with a DNA construct of thepresent invention is accomplished by well known methods that typicallydepend on the type of vector used. With regard to transformation ofprokaryotic host cells, see, for example, Cohen et al (1972) Proc. Natl.Acad. Sci. USA 69, 2110 and Sambrook et al (1989) Molecular Cloning, ALaboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor,N.Y. Transformation of yeast cells is described in Sherman et al (1986)Methods In Yeast Genetics, A Laboratory Manual, Cold Spring Harbor, N.Y.The method of Beggs (1978) Nature 275, 104-109 is also useful. Withregard to vertebrate cells, reagents useful in transfecting such cells,for example calcium phosphate and DEAE-dextran or liposome formulations,are available from Stratagene Cloning Systems, or Life TechnologiesInc., Gaithersburg, Md. 20877, USA.

Electroporation is also useful for transforming cells and is well knownin the art for transforming yeast cell, bacterial cells and vertebratecells.

For example, many bacterial species may be transformed by the methodsdescribed in Luchansky et al (1988) Mol. Microbiol. 2, 637-646incorporated herein by reference. The greatest number of transformantsis consistently recovered following electroporation of the DNA-cellmixture suspended in 2.5×PEB using 6250V per cm at 251 μFD.

Methods for transformation of yeast by electroporation are disclosed inBecker & Guarente (1990) Methods Enzymol. 194, 182.

Successfully transformed cells, ie cells that contain a DNA construct ofthe present invention, can be identified by well known techniques. Forexample, cells resulting from the introduction of an expressionconstruct of the present invention can be grown to produce thepolypeptide of the invention. Cells can be harvested and lysed and theirDNA content examined for the presence of the DNA using a method such asthat described by Southern (1975) J. Mol. Biol. 98, 503 or Berent et al(1985) Biotech. 3, 208. Alternatively, the presence of the protein inthe supernatant can be detected using antibodies as described below.

In addition to directly assaying for the presence of recombinant DNA,successful transformation can be confirmed by well known immunologicalmethods when the recombinant DNA is capable of directing the expressionof the protein. For example, cells successfully transformed with anexpression vector produce proteins displaying appropriate antigenicity.Samples of cells suspected of being transformed are harvested andassayed for the protein using suitable antibodies.

Thus, in addition to the transformed host cells themselves, the presentinvention also contemplates a culture of those cells, preferably amonoclonal (clonally homogeneous) culture, or a culture derived from amonoclonal culture, in a nutrient medium.

In another aspect the present invention provides antibodies to PP1cwhich act in an analogous manner to the peptides of interest. Antibodiesto the peptides themselves are also provided and these may themselves beused to affect cell metabolism or function.

Peptides in which one or more of the amino acid residues are chemicallymodified, before or after the peptide is synthesised, may be usedproviding that the function of the peptide, namely the production ofspecific antibodies in vivo, remains substantially unchanged. Suchmodifications include forming salts with acids or bases, especiallyphysiologically acceptable organic or inorganic acids and bases, formingan ester or amide of a terminal carboxyl group, and attaching amino acidprotecting groups such as N-t-butoxycarbonyl. Such modifications mayprotect the peptide from in vivo metabolism. The peptides may be presentas single copies or as multiples, for example tandem repeats. Suchtandem or multiple repeats may be sufficiently antigenic themselves toobviate the use of a carrier. It may be advantageous for the peptide tobe formed as a loop, with the N-terminal and C-terminal ends joinedtogether, or to add one or more Cys residues to an end to increaseantigenicity and/or to allow disulphide bonds to be formed. If thepeptide is covalently linked to a carrier, preferably a polypeptide,then the arrangement is preferably such that the peptide of theinvention forms a loop.

According to current immunological theories, a carrier function shouldbe present in any immunogenic formulation in order to stimulate, orenhance stimulation of, the immune system. It is thought that the bestcarriers embody (or, together with the antigen, create) a T-cellepitope. The peptides may be associated, for example by cross-linking,with a separate carrier, such as serum albumins, myoglobins, bacterialtoxoids and keyhole limpet haemocyanin. More recently developed carrierswhich induce T-cell help in the immune response include the hepatitis-Bcore antigen (also called the nucleocapsid protein), presumed T-cellepitopes such as Thr-Ala-Ser-Gly-Val-Ala-Glu-Thr-Thr-Asn-Cys (SEQ ID No1), beta-galactosidase and the 163-171 peptide of interleukin-1. Thelatter compound may variously be regarded as a carrier or as an adjuvantor as both. Alternatively, several copies of the same or differentpeptides of the invention may be cross-linked to one another; in thissituation there is no separate carrier as such, but a carrier functionmay be provided by such cross-linking. Suitable cross-linking agentsinclude those listed as such in the Sigma and Pierce catalogues, forexample glutaraldehyde, carbodiimide and succinimidyl4-(N-maleimidomethyl)cyclohexane-1-carboxylate, the latter agentexploiting the —SH group on the C-terminal cysteine residue (ifpresent).

If the peptide is prepared by expression of a suitable nucleotidesequence in a suitable host, then it may be advantageous to express thepeptide as a fusion product with a peptide sequence which acts as acarrier. Kabigen's “Ecosec” system is an example of such an arrangement.

The peptide of the invention may be linked to other antigens to providea dual effect.

In a yet further aspect the present invention provides a method ofdiagnosis of abnormalities of cellular metabolism, said methodcomprising analysing the naturally occurring peptide(s) or thenucleotide sequences encoding therefore and comparing the results to thepeptides described herein.

The peptides of the present invention may also be used in diagnosis andthis aspect is also covered by the present invention.

The specificity of the catalytic subunit of protein phosphatase-1 (PP1c)is modified by regulatory subunits that target it to particularsubcellular locations. For the first time we have identifiedPP1c-binding domains on G_(L) and G_(M), the subunits that target PP1cto hepatic and muscle glycogen, respectively, and on M₁₁₀, the subunitthat targets PP1c to smooth muscle myosin. The peptide G_(M)-(G63-T93)interacted with PP1c and prevented G_(L) from suppressing thedephosphorylation of glycogen phosphorylase, but it did not dissociateG_(L) from PP1c or affect other characteristic properties of thePP1_(GL) complex. These results indicate that G_(L) contains twoPP1c-binding sites, the region which suppresses the dephosphorylation ofglycogen phosphorylase being distinct from that which enhances thedephosphorylation of glycogen synthase. At higher concentrations,G_(M)-(G63-N75) had the same effect as G_(M)-(G63-T93), but not if Ser67was phosphorylated by cyclic AMP-dependent protein kinase. Thusphosphorylation of Ser67 dissociates G_(M) from PP1c because phosphateis inserted into the PP1c-binding domain of G_(M). The fragmentsM₁₁₀-(M1-E309) and M₁₁₀-(M1-F38), but not M₁₁₀-(D39-E309), mimicked theM₁₁₀ subunit in stimulating dephosphorylation of the smooth musclemyosin P-light chain and heavy meromyosin in vitro. However, in contrastto the M₁₁₀ subunit and M₁₁₀-(M1-E309), neither M₁₁₀-(M1-F38) norM₁₁₀-(D39-E309) suppressed the PP1c-catalysed dephosphorylation ofglycogen phosphorylase. These observations suggest that the region whichstimulates the dephosphorylation of myosin is situated within theN-terminal 38 residues of the M₁₁₀ subunit, while the region whichsuppresses the dephosphorylation of glycogen phosphorylase requires thepresence of at least part of the region 39-296 which contains sevenankyrin repeats. M₁₁₀-(M1-F38) displaced G_(L) from PP1c, whileG_(M)-(G63-T93) displaced M₁₁₀ from PP1c in vitro. These observationsindicate that the region(s) of PP1c that interact with G_(M)/G_(L) andM₁₁₀ overlap, explaining why different forms of PP1c contain just asingle targeting subunit.

We also disclose the structure of PP1c in complex with a portion of atargeting subunit, and show that changing key amino acid residues in thesubunit disrupts its interaction with PP1c. These studies identify acritical structural motif in targeting subunits involved in theinteraction with PP1c as well as the recognition site on PP1c itself.These findings will facilitate the rational design of agents such aspeptides or other forms of small cell-permeant molecules that act bydisrupting PP1-targeting subunit interactions. Given the structuralmotif and the coordinates of the atoms in the crystal structure, it iswithin the scope of the abilities of a skilled molecular modeller toproduce small cell-permeant molecules, which can enter cells naturally,and possess either the same motif, or an analogous structure to give thesame functional properties to the molecule. Thus the small cell-permeantmolecule can have a precise copy of the motif, or one which isfunctionally equivalent. The molecule can be a peptide, but other typesof molecules, which are transferred across the plasma membrane of cells,may be preferred.

Several mammalian PP1c targeting subunits have been isolated andcharacterised, including the G_(M) subunit that targets PP1c to both theglycogen particles and sarcoplasmic reticulum of striated muscles (Tanget al., 1991; Chen et al., 1994), the G_(L) subunit that targets PP1c toliver glycogen (Moorhead et al., 1995; Doherty et al., 1995), the M₁₁₀subunits responsible for the association of PP1c with the myofibrils ofskeletal muscle (Moorhead et al., 1994; Alessi et al., 1992) and smoothmuscle (Alessi et al., 1992; Chen et al., 1994), the p53 binding proteinp53BP2 (Helps et al., 1995) and the nuclear protein NIPP-1 (Jagiello etal., 1995; Van Eynde et al., 1995). PP1c is also reported to interactwith other mammalian proteins such as the retinoblastoma gene product(Durphee et al., 1993), an RNA splicing factor (Hirano et al., 1996),ribosomal proteins L5 (Hirano et al., 1995) and RIPP-1 (Beullens et al.,1996), a 110 kDa nuclear protein yet to be identified (Jagiello et al.,1995), kinesin-like proteins and small cytosolic proteins, inhibitor-1,DARPP-32 and inhibitor-2 (Cohen, 1989; Cohen, 1992, Hubbard and Cohen,1993). Moreover, a number of distinct PP1-regulatory subunits have beenidentified in yeast (reviewed by Stark, 1996). We attempted to identifywhich regions of the G_(M) and M₁₁₀ subunits were involved in binding toPP1c. These studies led to the identification of relatively smallpeptides from each targeting subunit that were capable of interactingwith PP1c. Peptides comprising residues 63-93, 63-80 and 63-75 of G_(M)bound to PP1c, dissociating it from G_(L), while the N-terminal 38residues of the M₁₁₀ subunit (M₁₁₀[1-38]) mimicked the intact M₁₁₀subunit in enhancing the rate at which PP1c dephosphorylated the 20 kDamyosin light chain (MLC₂₀) subunit of smooth muscle myosin (Johnson etal., 1996).

The present invention thus provides peptides comprising the N-terminal38 residues of the M₁₁₀ subunit, and those comprising residues 63-93,63-80 and 63-75 of G_(M).

Phosphorylation of Ser 67 of G_(M) by protein kinase A (PKA) disruptsthe interaction of G_(M) with PP1c (Dent et al., 1990) and a similardisruption is also observed following the phosphorylation of Ser 67 ofthe G_(M)[63-75] peptide (Johnson et al., 1996). The finding thatG_(M)[63-93] disrupted the interaction between PP1c and the M₁₁₀subunit, and prevented M₁₁₀ from enhancing the MLC₂₀ phosphataseactivity of PP1c implies that the binding of M₁₁₀ and G_(M) to PP1c aremutually exclusive.

Thus the invention contemplates the substitution or modification of anamino acid in any such peptide.

To understand the basis for the recognition by PP1c of regulatorysubunits, and peptides derived from these subunits, we co-crystallised acomplex of PP1c with the G_(M)[63-75] peptide and determined thestructure at 3.0 Å resolution. These experiments have demonstrated thatresidues 64 to 69 of the peptide are bound in an extended conformationto a hydrophobic channel within the C-terminal region of PP1c. Theresidues in G_(M)[63-75] that interact with PP1c lie in anArg/Lys-Val/Ile-Xaa-Phe motif common to M₁₁₀[1-38] and almost all knownmammalian PP1-binding proteins. Substituting Val or Phe by Ala in theG_(M)[163-75] peptide, and deleting the VxF motif from the M₁₁₀[1-38]peptide, abolished the ability of both peptides to interact with PP1c.These findings identify a recognition site on PP1c for a criticalstructural motif involved in the interaction of targeting subunits withPP1.

Particularly preferred peptides are derived from residues 63 to 69 ofG_(M) and comprise the motif Arg/Lys-Val/Ile-Xaa-Phe. Peptides derivedfrom M₁₁₀ (or any other source) and also including the motif are alsoincluded in the scope of the invention.

Preferred peptides may also be substantially or wholly made up ofhydrophobic residues.

The identification of this area of PP1c necessary for binding to thevarious subunits allows the design of agents to specifically disrupt theinteraction at this area. Such disruption may, for example, increase thephosphorylation of the protein phospholamban in cardiac muscle and thusincrease the force and rate of contraction of the muscle. This providesa possible treatment for congestive heart failure. Also, the specificdisruption of the complex of PP1 and p53BP2 may prevent PP1 fromdephosphorylating the tumour suppressor protein p53, thus enhancingphosphorylation of p53, its ability to bind to DNA, and thus its abilityto act as a tumour suppressor.

The identification of the key motif in targetting subunits that bind toPP1 also provides the means to produce targetting subunits that can nolonger interact with PP1. Over-expression of these mutant targettingsubunits provides a powerful new way to determine the functions ofdifferent targetting subunits in vivo.

Abbreviations

-   -   PP1, protein phosphatase-1    -   PP1c, catalytic subunit of PP1    -   PP1, —isoform of PP1c    -   PP1_(G), glycogen-associated form of PP1    -   PP1_(M), myosin-associated form of PP1    -   G_(M), glycogen-binding subunit of PP1 from striated muscle    -   G_(L), glycogen-binding subunit of PP1 from liver    -   NIPP1, nuclear inhibitor of PP1    -   DARPP, dopamine and cyclic AMP-regulated phosphoprotein    -   M₂₁ and M₁₁₀, myofibrillar-binding subunits of PP1 with        molecular masses of 21 kDa and 110 kDa respectively.    -   PKA, cyclic AMP-dependent protein kinase    -   PhMeSO₂F, phenylmethylsuphonyl fluoride    -   GST, glutathione-S-transferase    -   MLC₂₀, myosin light chain of molecular mass 20 kDa.

The invention is now described in more detail by reference to thefollowing Examples and Figures wherein:

FIG. 1 shows that the N-terminal 118 residues of human G_(M) interactwith PP1c.

GST-G_(M) fusion proteins were electrophoresed on 10% SDS/polyacrylamidegels and stained with Coomassie blue (lanes 1-3) or, after transfer tonitrocellulose, probed with digoxygenin-labelled PP1γ (lanes 4-6) as in[9]. Lanes 1 and 4, GST-G_(M)-(E2-D118); Lanes 2 and 5,GST-G_(M)-(H100-P350); Lanes 3 and 6, GST. The positions of the markerproteins glycogen phosphorylase (97 kDa), bovine serum albumin (66 kDa),ovalbumin (43 kDa), carbonic anhydrase (29 kDa) and soybean trypsininhibitor (20 kDa) are indicated.

FIG. 2 shows that synthetic peptides between residues 63 and 93 ofrabbit G_(M) stimulate the phosphorylase phosphatase activity ofPP1_(GL).

Hepatic glycogen particles were diluted in assay buffer to 0.6phosphorylase phosphatase (PhP) mU per ml, incubated for 15 minutes at30° C. with G_(M)-(G63-T93) (closed circles), G_(M)-(G63-K80) (opencircles) or G_(M)-(G63-N75) (closed triangles) and assayed as describedin Example 1. The open triangles show the effect of G_(M)-(G63-N75)which had been phosphorylated at Ser67 by PKA (pG_(M)-(G63-N75)).Similar results were obtained in four experiments.

FIG. 3 shows that removal of the M₂₁ subunit from smooth muscle PP1_(M)does not affect its MLC20 phosphatase:phosphorylase phosphatase activityratio.

(A) Purified smooth muscle PP1_(M) was electrophoresed on a 12%SDS/polyacrylamide gel, and either stained with Coomassie blue (lane 1)or immunoblotted [32] with antibodies specific for the M₂₁ subunit (lane2) or the M₁₁₀ subunit (lane 3). The positions of the M₁₁₀ subunit, theM₂₁ subunit and PP1c are marked.

(B) Purified PP1_(M) (lane 1) or PP1_(M) lacking the M₂₁ subunit (lane2) were electrophoresed on a 12% SDS polyacrylamide gel, transferred tonitrocellulose and immunoblotted with mixed, affinity-purifiedantibodies to the M₁₁₀ and M₂₁ subunits. The M₁₁₀ and M₂₁ subunits aremarked. The activity ratio, MLC₂₀ phosphatase (MP):phosphorylasephosphatase (PhP) of the two preparations is also shown. Similar resultswere obtained in three different experiments. The activity ratio MP:PhPof PP1c is 0.07.

FIG. 4 shows expressed fragments of the M₁₁₀ subunit before and aftercleavage of the GST-fusion proteins with thrombin.

Purified GST-fusion proteins were electrophoresed on a 15%SDS/polyacrylamide gel and stained with Coomassie blue. Lane 1,GST-M₁₁₀-(M1-A150); Lane 2, GST-M₁₁₀-(D39-E309); Lane 3,GST-M₁₁₀-(M1-E309); Lane 4, GST-M₁₁₀-(L24-Y496). Lanes 5-8 are the sameas Lanes 1-4 except that the fusion proteins were cleaved with thrombin.The positions of the marker proteins glycogen phosphorylase (97 kDa),bovine serum albumin (66 kDa), ovalbumin (43 kDa), carbonic anhydrase(29 kDa), GST (26 kDa) and soybean trypsin inhibitor (20 kDa) aremarked.

FIG. 5 shows the effect of M₁₁₀ subunit fragments on PP1c-catalyseddephosphorylation of MLC₂₀ and glycogen phosphorylase.

A,B; Effects of M₁₁₀-(M1-E309) (closed circles), M₁₁₀-(M1-F38) (opencircles) and M₁₁₀-(D39-E309) (open triangles) on the MLC₂₀ phosphatase(B) and phosphorylase phosphatase (B) activities of PP1c were measuredafter incubating PP1c for 15 minutes at 30° C. with each fragment. Theresults are presented as a percentage of those obtained in experimentswhere the M₁₁₀ fragments were omitted.

C,D; The effect of M₁₁₀-(M1-A150) (open circles) and M₁₁₀-(L24-Y496)(closed circles) on the MLC₂₀ phosphatase (C) and phosphorylasephosphatase (D) activities of PP1c were measured as in A,B.

FIG. 6 shows the effect of M₁₁₀-(M1-F38) and M₁₁₀-(M1-E309) on thedephosphorylation of glycogen synthase by PP1c.

The glycogen synthase phosphatase activity of PP1c was measured after a15 minute incubation at 30° C. with the indicated concentrations ofM₁₁₀-(M1-F38) and M₁₁₀-(M1-E309). Similar results were obtained in threedifferent experiments.

FIG. 7 shows that G_(M)-(G63-T93) dissociates PP1_(M).

(A) The phosphorylase phosphatase (PhP) activity of PP1_(M) (closedcircles) and its MLC₂₀ phosphatase (MLCP) activity (open circles) wereassayed after preincubation for 15 minutes at 30° C. with the indicatedconcentrations of G_(M)-(G63-T93). Activities are shown relative tocontrol incubations in which G_(M)-(G63-T93) was omitted. Similarresults were obtained in three experiments.

(B,C) PPIM was incubated for 15 minutes at 30° C. in the absence (B) andpresence (C) of 10 M G_(M)-(G63-T93), then passed through a 30×1 cmcolumn of Superose 12 equilibrated at ambient temperature in 50 mMTris/HCl pH 7.5, 0.2M NaCl, 0.1 mM EGTA, 0.1% (by vol)2-mercaptoethanol, 0.03% (by mass) Brij 35 in the absence (B) orpresence (C) of 1 μM G_(M)-(G63-T93). Fractions (0.25 ml) were assayedfor MLC₂₀ phosphatase (MLCP) in B and for phosphorylase phosphatase(PhP) activity in C. The arrows denote the position of ferritin (450kDa) and ovalbumin (43 kDa).

FIG. 8 shows that G_(M)-(G63-T93) prevents M₁₁₀-(M1-F38) orM₁₁₀-(M1-E309) from modulating the substrate specificity of PP1c.

(A) The MLC₂₀ phosphatase activity of PP1c was assayed after incubationfor 15 minutes at 30° C. in the presence or absence of 1 μMG_(M)-(G63-T93) and either 0.1 μM M₁₁₀-(M1-F38) or 0.1 nMM₁₁₀-(M1-E309).

(B) The phosphorylase phosphatase activity of PP1c was assayed as in Ain the presence or absence of 1 μM G_(M)-(G63-T93) and 1.0 nMM₁₁₀-(M1-E309). The results are presented (SEM for three experiments) asa percentage of the PP1c activity measured in the absence ofG_(M)-(63-T93), M₁₁₀-(M1-F38) or M₁₁₀-(M1-E309).

FIG. 9 shows the location of PP1c-binding domains on the G_(M) and M₁₁₀targeting subunits and their effects on PP1 activity.

The hatched boxes in the M₁₁₀ subunit denote the positions of theankyrin repeats.

FIG. 10 shows a stereo view of the electron density corresponding to thepeptide. A: Initial 2-fold averaged electron map. B: map calculatedusing 3Fo-2Fc coefficients and phases calculated from the final refinedmodel. Displayed using TURBO-FRODO.

FIG. 11 shows the structure of PP1-G_(M)[63-75] peptide complex. A.Stereo view of a ribbons diagram of PP1c to indicate the position of thepeptide binding channel at the interface of the two β-sheets of theβ-sandwich. The peptide atoms are represented as ball-and-stick(MOLSCRIPT, Kraulis, 1991).

B. View of the surface of PP1c to show the hydrophobic peptide bindingchannel. Residues 63′ to 69′ (GRRVSFA) (SEQ ID No 2) of the G_(M)[63 75]peptide are shown as sticks. Drawn with TURBO-FRODO.

C. Stereo view of the G_(M)[63-75] peptide at the recognition site ofPP1 to indicate polar interactions between peptide and protein and theformation of the β-sheet between Ser 67′—Ala 69′ and 14 of PP1. Drawnwith TURBO-FRODO.

E. Details of the structure of the peptide binding site to showhydrophobic interactions between PP1c and Val 66′, Phe 68′ and Ala 69′of the G_(M)[68-75] peptide (MOLSCRIPT, Kraulis, 1991).

FIG. 12 shows a sequence alignment of PP1-regulatory subunits in thevicinity of the (R/K)(V/I) x F motif. (A) mammalian PP1-bindingsubunits. G_(M), Tang et al., 1991; G L, Docherty et al., 1995;G_(L)-related protein, Doherty et al., 1996; p53BP2, Helps et al., 1995;NIPP-1, Bollen et al., 1995; splicing factor PSF, Hirano et al., 1996;M₁₁₀ subunit, Chen et al., 1994; inhibitor-1, Aitken et al., 1982;DARPP-32, Williams et al., 1986. (B) PP1-binding proteins in S.cerevisiae. GAC1 (Francois et al., 1992); PIG2 GIP1, GIP2, YIL045W (Tuet al., 1996); REG1, REG2 (Tu and Carlson, 1995; Frederick and Tatchell,1996); SCD5 (Nelson et al 1996; Tu et al 1996). The region homologous tothe RRVSFA (SEQ ID No 3) motif in G_(M) which intersects with PP1c isboxed.

FIG. 13 shows the disruption of the interactions between PP1c and theG_(L) and M₁₁₀ subunits by a synthetic peptide from p53BP2. (A) PP1_(M)from chicken gizzard smooth muscle (Alessi et al., 1992) was diluted andincubated for 15 min at 30° C. with the peptideGKRTNLRKTGSERIAHGMRVKFNPLALLLDSC (SEQ ID No 4), corresponding to thesequence in p53BP2 that contains the RVXF motif. Reactions were startedwith either ³²P-labelled MLC₂₀ or glycogen phosphorylase and the MLC₂₀phosphatase (open circles) and phosphorylase phosphatase (PhP, closedcircles) activities were determined. The results are expressed as apercentage of the activity determined in control incubations where thep53BP2 peptide was omitted (100%). Similar results were obtained inthree separate experiments. (B) same as (A) except that the peptide wasincubated with diluted hepatic glycogen particles containing PP1-G_(L)before measuring the PhP activity. Similar results were obtained inthree separate experiments.

FIG. 14 shows the effect of M₁₁₀[M1-F38] and M₁₁₀[M1-K35] on thePP1c-catalysed dephosphorylation of MLC₂₀ M₁₁₀[M1-F38] (1-38, opencircles) or M₁₁₀[M1-K35] (1-35, closed circles) were incubated with PP1cfor 15 min at 30° C. and reactions started with the ³²P-labelled MLC₂₀substrate. The results are expressed as a % of the activity determinedin control incubations where the M₁₁₀ peptides were omitted (100%).Similar results were obtained in three separate experiments.

FIG. 15 shows the effect of synthetic peptides derived from the M₁₀₀ andG_(M) subunits on the phosphorylase phosphatase activity of PP1-G_(L).(A) Hepatic glycogen protein particles containing PP1-G_(L) were dilutedand incubated for 15 min at 30° C. with the indicated concentrations ofeither M₁₁₀[M1-F38] (open circles) or M₁₁₀[M1-K35] (closed circles) andthe phosphatase reactions were initiated by addition of ³²P-labelledglycogen phosphorylase. The results are expressed as a percentage of theactivity determined in control incubations where the M₁₁₀ peptides wereomitted. Similar results were obtained in three separate experiments.(B) The experiment was carried out as in (A), except that the peptideG_(M)[G63-N75] (“wild type”, WT) and variants in which either Val 66(V66A) (closed triangles) or Phe 68 (F68A) (closed circles) were changedto Ala, were used instead of the M₁₁₀ peptides. Similar results wereobtained in three separate experiments.

FIG. 16 shows a stereo view of a ribbons diagram of a model ofPP1-phospho-inhibitor-1 complex. The side chains of Ile 10, Phe 12 andpThr 35 of phospho-inhibitor-1 are shown with the main-chain atoms ofresidues 8 to 36 of the inhibitor indicated as a shaded ribbon. Drawnwith MOLSCRIPT (Kraulis 1991).

FIG. 17 shows a comparison of rat and chicken gizzard M₁₁₀ and M₂₁subunits (SEQ ID NOs:29 to 31).

Vertical lines indicate identical residues, colons denote similarresidues in the rat and chicken M₁₁₀ sequences and deletions are shownby dots. (A) Comparison of M₁₁₀ subunits. Underlined residues in the ratM₁₁₀ subunit (Rat1) are deleted in some rat aorta forms and underlinedresidues in the chicken M₁₁₀ subunit (Ch1) are deleted in some chickengizzard forms [5, 8]. Dashed lines above residues indicate amino acidsdeleted in the rat kidney M₁₁₀ subunit [9. The alternative C-terminalsequences of rat uterus M₁₁₀ subunit are shown as Rat1 and Rat2. Leucineresidues in the C-terminal leucine zipper motif are double underlined.(B). The C-terminal sequence of the M₁₁₀ subunit is structurally relatedto the M₂₁ subunit. The sequence of the chicken M₂₁ subunit [5] iscompared with the C-terminal sequences of Rat2 and Ch1 from A.Identities between Ch1 and Rat2 are shown in boldface type.

FIG. 18 shows immunoprecipitation and immunoblotting of PP1_(M) inextracts from chicken gizzard myofibrils.

A. Antibodies specific for the M₁₁₀ and/or M₂₁ subunitsimmunoprecipitate most of the myosin P-light chain phosphatase activityin myofibrillar extracts. PP1_(M) was immunoprecipitated with eithercontrol IgG, antibody raised against the PP1_(M) holoenzyme, antibodyspecific for the M₁₁₀ subunit or antibody specific for the M₂₁ subunit,as described under Methods in Example 3. The figure shows activitypresent in the supernatant (S, open bars) or pellet (P, filled bars) asa percentage of that measured before centrifugation. The results shownare the average (±S.E.M.) for three separate experiments each assayed induplicate. B, The M₁₁₀ and M₂₁ subunits are present in similar molarproportions in myofibrillar extracts and in purified PP1_(M). 10 ng(track 1) or 3 ng (track 3) of purified PP1_(M) or 12 μg (track 2) or3.6 μg (track 4) of myofibrillar extract was electrophoresed on a 12%SDS/polyacrylamide gel, transferred to nitrocellulose and immunoblottedwith mixed affinity-purified antibodies to the M₁₁₀ and M₂₁ subunits asin [22]. The positions of the two subunits are marked. The resultsindicate that PP1_(M) comprises about 0.1% of the myofibrillar protein.

FIG. 19 shows the identification of the region on the M₁₁₀ subunit thatinteracts with the M₂₁ subunit.

A) PP1_(M) 5 μg (track 1), 10 μg bacterial extract containingM₁₁₀-(R714-I1004) (track 2), MBP-M₁₁₀-(R714-I004) 1 μg (track 3),MBP-M₁₁₀-(R714-L934) 1 μg (track 4), MBP-M₁₁₀-(K933-I1004) 1 μg (track5), MBP 1 μg (track 6), M₁₁₀-(M1-E309) 2 μg (lane 7) and M₁₁₀-(M1-S477)2 μg (track 8) were run on a 12% SDS/polyacrylamide gel and stained withCoomassie Blue. B) same as A) except that 10-fold less protein waselectrophoresed and after transfer to nitrocellulose the proteins wereprobed with digoxigenin-labelled M₂₁ subunit (0.2 μg/ml). C) same as B)except that, after electrophoresis, the proteins were transferred tonitrocellulose and probed with digoxigenin-labelled M₂₁-(M1-L146) (0.2μg/ml).

FIG. 20 shows the identification of the region of the M₂₁ subunitinvolved in interaction with the M₁₁₀ subunit and in dimerization.

A) GST-M₂₁ 5 μg (track 1), M₂₁ 5 μg (track 2), M₂₁-(M1-L146) 5 μg (track3), M₂₁-(M1-E110) 20 μg (track 4) and M₂₁-(E110-K186) 5 μg (track 5)were run on 16.5% polyacrylamide gels and stained with Coomassie Blue.The marker proteins ovalbumin (43 kDa) and carbonic anhydrase (29 kDa)are indicated. B) GST-M₂₁ 0.5 μg (track 1), M₂₁ 0.5 μg (track 2),M₂₁-M1-L146) 0.5 μg (track 3), M₂₁-(M1-E110) 5 μg (track 4) andM₂₁-(E110-K186) 5 μg (track 5) were electrophoresed as in A) and aftertransfer to nitrocellulose the blots were probed withdigoxigenin-labelled MBP-M₁₁₀-(K933-I1004) (0.2 μg/Ml). C) same as B)except that, after electrophoresis, the proteins were transferred tonitrocellulose and probed with digoxigenin-labelled M₂₁ subunit (0.2μg/ml).

FIG. 21 shows that the M₂₁ subunit and M₂₁-(M1-L146) interact with theM₁₁₀ subunit and themselves, but not with PP1.

PP1_(M) (0.5 μAg) was electrophoresed on a 12% SDS/polyacrylamide gel,transferred to nitrocellulose and probed with digoxigenin-labelled M₂₁subunit (0.2 μg/ml) (track 1) or digoxigenin-labelled M₂₁-(M1-L146) (0.2μg/ml) (track 2). The positions of the M₁₁₀ subunit, the M₂₁ subunit andPP1c are marked.

FIG. 22 shows that removal of the M₂₁ subunit from smooth muscle PP1_(M)does not prevent it from being pelleted with myosin.

The PP1 catalytic subunit (PP1c), PP1_(M), or PP1_(M) lacking the M₂₁subunit, PP1_(M)(ΔM₂₁), each at 30 nM, were incubated for 15 min at 0°C. with 1 μM myosin and centrifuged (see Methods of Example 3). Thefigure shows the myosin P-light chain phosphatase activity present inthe supernatant (S, open bars) or pellet (P, filled bars) as apercentage of that measured before centrifugation. The results shown arethe average (±S.E.M.) for three separate experiments each assayed induplicate.

FIG. 23 shows the identification of a region of the M₁₁₀ subunit whichbinds to myosin.

(A); PP1_(M), M₁₁₀-(M1-S477) and GST-M₁₁₀-(M377-K976), each at 30 nMwere incubated for 15 min at 0° C. with 1 μM myosin and centrifuged. Thesupernatants (S), resuspended pellets (P) and the suspension beforecentrifugation (T, total) were electrophoresed on 12% SDS/polyacrylamidegels, transferred to nitrocellulose and immunoblotted with antibodiesraised against the PP1_(M) holoenzyme. No protein was pelleted in theabsence of myosin (not shown). The positions of the marker proteinsmyosin heavy chain (200 kDa), glycogen phosphorylase (97 kDa), bovineserum albumin (66 kDa), ovalbumin (43 kDa), carbonic anhydrase (29 kDa)and soybean trypsin inhibitor (20 kDa) are indicated. (B) Theexperiments were carried out as in (A), except that the M₁₁₀ fragmentsand M₂₁ subunit were used at 100 nM, the 8.5 kDa M₁₁₀-(K933-I1004)fragment was electrophoresed on a 16.5% polyacrylamide gel andimmunoblotting was carried out with affinity purified antibodies (seeMethods). A small amount of M₁₁₀-(R714-I1004) pelleted in the absence ofmyosin. This was probably due to aggregation in the bacterial extractsince this did not happen when it was complexed to the M₂₁ subunit (datanot shown). No other protein was pelleted in the absence of myosin.

FIG. 24 shows that the isolated M₂₁ subunit binds to myosin.

(A); Myosin (1 μM) was mixed with 50 μM, 20 μM or 10 μM M₂₁ subunit togive the molar ratios M₂₁:myosin dimer indicated. After 15 min at 0° C.,the solutions were centrifuged and the supernatants (S), resuspendedpellets (P) and the suspension before centrifugation (T, total) wereelectrophoresed on 12% SDS/polyacrylamide gels and stained withCoomassie blue. The positions of the myosin heavy chain (MHC) and theM₂₁ subunit are indicated. The myosin light chains migrate faster thanthe M₂₁ subunit and are not visible at these loadings.

(B); Myosin (track A) was purified from chicken gizzard, and the myosin“rod” domain (track B) and light meromyosin (track C) produced bydigestion of myosin with papain and chymotrypsin, respectively. Thesethree proteins, all at 1 μM, were then mixed with M₂₁ subunit (track D)to give a molar ratio M₂₁:myosin dimer of 10:1 and, after 15 min at 0°C., the solutions were centrifuged and the supernatants (S), resuspendedpellets (P) and the suspension before centrifugation (T, total) wereelectrophoresed on 12% SDS/polyacrylamide gels and stained withCoomassie blue. The slightly faster migrating band in the M₂₁ subunitpreparation was shown by amino acid sequencing to be N-terminallytruncated commencing at residue 16. (C); same as (B), except thatM₂₁-(M1-L146) (track D) replaced the M₂₁ subunit.

FIG. 25 gives a schematic representation of the regions on the M₁₁₀subunit from chicken gizzard that interact with PP1c, myosin and the M₂₁subunit.

PP1c binds to the KVKF (SEQ ID No 5) motif between residue 35 and 38,just N-terminal to the seven ankyrin repeats (hatched vertical lines)that suppress the dephosphorylation of substrates other than myosin.Residues 1-38 of the M₁₁₀ subunit enhance the dephosphorylation ofmyosin. The M₂₁ subunit binds to the C-terminal 72 residues of the M₁₁₀subunit which are 43% identical in amino acid sequence to residues87-161 of the M₂₁ subunit. The dephosphorylated form of myosin binds toM₁₁₀-(R714-I1004) but not to M₁₁₀-(K933-I1004), suggesting that myosinbinds N-terminal to the M₂₁ subunit.

EXAMPLE 1 Identification of Protein Phosphatase 1-binding Domains on theGlycogen and Myofibrillar Targeting Subunits

Materials and Methods

Materials.

The myosin-associated form of PP1 (PP1_(M)) was from chicken gizzard [9]and the glycogen-associated form of PP1 (PP1_(G)) from rabbit skeletalmuscle [2]. The β isoform of PP1c was released from PP1_(G) byincubation for 2 hours in 2M LiBr, then purified by gel-filtration on a30×1 cm column of Superose 12 (Pharmacia, Milton Keynes, U.K.) in thepresence of 0.5M LiBr. Glycogen protein particles from rat liver [22]were used as the source of hepatic PP1_(G) Digoxygenin-labelled PP1c(γ₁-isoform, hereafter termed PP1) was prepared as in [9]. G_(L) wasexpressed in E. coli as a glutathione-S-transferase (GST) fusion protein[7], termed GST-G_(L). The catalytic subunit of PP2A from bovine heart(PP2AC) was provided by Dr R. MacKintosh in this Unit. Thephosphorylatable myosin light chain (MLC₂₀) and heavy meromyosin fromchicken gizzard were a gift from Dr M. Ikebe (Case Western ReserveUniversity, Cleveland, USA). Thrombin and benzamidine-Agarose werepurchased from Sigma (Poole, UK).

Peptide Synthesis.

Peptides were synthesised on an Applied Biosystems 430A peptidesynthesiser and their purity and concentration established by highperformance liquid chromatography, mass spectrometry and amino acidanalysis. The sequence of rabbit G_(M)-(G63-T93) isGRRVSFADNFGFNLVSVKEFDTWELPSVSTT (SEQ ID No 6) and the sequence ofM₁₁₀-(M1-F38) is MKMADAKQKRNEQLKRWIGSETDLEPPVVKRQKTKVKF (SEQ ID No 7).The peptide G_(M)-(G63-T93) was cleaved with Lys-C endoproteinase(Boehringer) and the peptide G_(M)-(E81-T93) thus generated was purifiedon a C₁₈ column. The peptides G_(M)-(G63-K80) and G_(M)-(G63-N75), weresynthesised, and the latter phosphorylated at Ser67 with the catalyticsubunit of cyclic AMP-dependent protein kinase (PKA), then bound to a 1ml C₁₈ column equilibrated in 0.1% (v/v) trifluoroacetic acid, washedwith 0.1% trifluoroacetic acid to remove excess ATP, eluted with 0.1%trifluoroacetic acid containing 70% acetonitrile, dried and dissolved inwater. The peptide G_(M)-(S40-Y55) was a gift from Dr Bruce Kemp (StVincent's Institute, Australia).

Preparation of Phosphorylated Proteins and Phospharase Assays.

³²P-labelled rabbit skeletal muscle phosphorylase a (containing 1.0 molphosphate per mol subunit) was prepared by phosphorylation withphosphorylase kinase [23], ³²P-labelled rabbit skeletal muscle glycogensynthase (containing 1.5 mol/mol subunit in the sites 3 region) wasprepared by phosphorylation with glycogen synthase kinase-3 [24]).³²P-labelled chicken gizzard MLC₂₀ and ³²P-labelled chicken gizzardheavy meromyosin (containing 1.0 mol phosphate per mol subunit) wereprepared by phosphorylation with smooth muscle myosin light chain kinase[9]. The dephosphorylation of phosphorylase a (10 μM), glycogen synthase(1 μM) and MLC₂₀ (1 μM) and heavy meromyosin (1 μM) was carried out asin [24]. One unit of activity (U) was that amount which released 1 moleof phosphate in one minute.

Construction of Vectors for the Expression of N-terminal Fragments ofthe G_(M) Subunit as Glutathione-S-transferase (GSZ) Fusion Proteins inE. coli.

G_(M)-(E2-RS75) was produced by inserting a SmaI-SmaI restrictionfragment, encoding amino acids 2-575 of human G_(M), from clone H1G11[5] into the Smal site of pGEX-KG (Pharmacia, Milton Keynes, U.K.). Thisresulted in the addition after residues 2-575 of amino acids EFPVVVVEF(SEQ ID No 8) before the stop codon. G_(M)-(E2-P243) was made bydeleting an NcoI-HindIII fragment of the G_(M)-(E²-R575) construct,resulting in termination after residue 243. G_(M)-(E2-D118), encodingamino acids 2-118, with a C-terminal addition of QLNSS was produced bydeleting a BglII-HindIII fragment of the G_(M)-E2-R575) construct.G_(M)-H100-P350) encoding amino acids 100-350 was made by inserting anEcoRI-HindIII digested PCR fragment prepared using primers 5′GCCGAATTCACACAGAAGAATATGTTTTAGCC 3′ (SEQ ID No 9) and 5′GCCGAAGCTTATGGAAAATTGACTGGATCTGTTG 3′ (SEQ ID No 10) into the same sitesof pGEX-KG. Restriction sites in the primers are underlined.

Construction of Vectors for the Expression of Tile Chicken Gizzard M₂₁Subunit in E. coli.

The entire coding region (M1-K186) of the M₂₁ subunit [10] was amplifiedby PCR using primers 5′ CGCGCATATGTCGTCGCTGTTCACCAGG 3′ (SEQ ID No 11)and 5′ GGCGGATCCCTACTTGGAGAGTTTGC 3′ (SEQ ID No 12), containingrestriction sites NdeI and BamHI (underlined). After cleavage with therestriction enzymes, the PCR fragment was cloned into the same sites ofthe bacterial expression vector pT7-7.

Production of Fragments of the Chicken Gizzard and Rat Aorta M₁₁₀subunits.

The C-terminal 291 residues M₁₁₀-(R714-I1004) of the chicken gizzardM₁₁₀ subunit were amplified by PCR using a primer 5′AGGAAGAATTCGTTCCACACGAAC 3′ (SEQ ID No 13) containing an EcoRIrestriction site (underlined) and a KS primer in the Bluescript vectorof the cDNA clone [10]. The EcoRI digested PCR fragment was subclonedinto the same site of pT7-7.

Rat aorta M₁₁₀ fragments were produced as GST-fusion proteins.M₁₁₀-(M1-A150) was amplified by PCR using primers A (5°CCTAGCCCGGGGATGAAGATGGCGGAC 3′) (SEQ ID No 14) and B (5′GCGGAAGCTTATGCTTCCTCCTCTGCAATATC 3′) (SEQ ID No 15), containing SmaI andHindIII restriction sites (underlined) and the SmaI-HindIII digested PCRfragment subcloned into the same sites of pGEX-KG. M₁₁₀-(M1-E309) wasproduced by subcloning a SmaI-HindIII digested PCR fragment amplifiedusing primers A and C (5′ CTAGAAGCTTCCATATTTGCTGTTGATTCAATC 3′) (SEQ IDNo 16) into the same sites of pGEX-KG. This resulted in one amino acid(A) being added after E309. M₁₁₀-(D39-E309) was produced by subcloning aSmaI-HindIII digested PCR fragment amplified using primers D(5′CCTAGCCCGGGGGACGATGGCGCCGTCTTCC 3′) (SEQ ID No 17) and C into thesame sites of pGEX-KG. An M₁₁₀-(L24-K976) was prepared by inserting aXhoI-XhoI restriction fragment of the entire M₁₁₀ cDNA in Bluescriptinto XhoI site of pGEX-KG, and M₁₁₀-(L24-Y496) expressed by deleting aNdeI-NdeI fragment of the L24-K976 construct and filling the overhangingends before ligating them. This resulted in the addition after Y496 ofamino acids MVAD (SEQ ID No 18) before the stop codon. The sequence ofall subclones produced after PCR amplification were verified using anApplied Biosystems 373A automated DNA sequencer and Taq dye terminatorcycle sequencing according to the manufacturer's instructions.

Expression of Proteins in E. coli.

All constructs were expressed in E. coli strain BL21(DE3)plysS. Cultureswere grown at 37° C. in Luria-Bertani medium in the presence of 100μg/ml ampicillin and 30 μg/ml chloramphenicol to an A600 of 0.4-0.6, andinduced with 50 μg/ml isopropylthiogalactoside for 8 hours at 25° C. orovernight at ambient temperature. After centrifugation for 10 minutes at7000×g (4° C.), cells from one liter of culture were resuspended in 20ml of 50 mM Tris-HCl pH 8.0, 0.1 M NaCl, 1 mM EDTA, 0. 1% (by vol)2-mercaptoethanol, 0.2 mM phenylmethylsulphonylfluoride (PhMeSO₂F), 1 mMbenzamidine (buffer A) and frozen at −80° C. After thawing, sodiumdeoxycholate (1 mg/ml ), 8 mM MgSO4 and 10 g/ml DNAase I were added, theextract incubated until it was no longer viscous, then made 6 mM inEDTA, 1 mM in benzamidine and 0.2 mM in PhMeSO₂F and centrifuged for 10minutes at 10,000×g. The soluble GST-fusion proteins were then purifiedfrom the supernatant by affinity chromatography on glutathione-Sepharose(Pharmacia).

The M₂₁ subunit and M₁₁₀(R714-I1004) C-terminal fragment from chickengizzard M₁₁₀ subunit, which were used for affinity purification of theanti-M₂₁ and anti-M₁₁₀ antibodies (see below) were obtained in inclusionbodies and therefore recovered in the pellets after centrifuging E. coliextracts at 10,000×g. M₁₁₀-(R714-I1004) was solubilised by resuspensionin Buffer A containing 0.5% (by mass) Triton X-100 and was >95% pure.The M₂₁ subunit was not solubilised by this procedure but, after washingthe pellets in 0.5% Triton X-100, was dissolved by sonication in 0.5%trifluoroacetic acid; its purity was about 20%.

M₁₁₀ GST-fusion proteins (1-9 mg/ml in 50 mM Tris/HCl, 2.5 mM CaCl2, 150mM NaCl and 0. 1% (by vol) 2-mercaptoethanol) were cleaved by incubationfor 20 minutes at 30° C. with 20 μg/ml thrombin. Benzamidine-Agarose(0.2 ml) was added and, after incubation (with rotation) for 30 minutesat ambient temperature, the benzamidine-Agarose containing the attachedthrombin was removed, and the supernatant dialysed against 50 mMTris-HCl pH 7.5, 0.1 mM EGTA, 0.1% (by vol) 2-mercaptoethanol, 10%glycerol and stored in aliquots at −80° C. After cleavage with thrombin,all fragments of the M₁₁₀ subunit, except M₁₁₀-(L24-Y496), commencedwith the sequence GSPG (SEQ ID No 19) before the initiating residue ofthe GST-fusion proteins. The M₁₁₀-(24-Y496) was preceded by the sequenceGSPGISGGGGGILDSMGR (SEQ ID No 20).

Production of Antibodies that Recognise the M₁₁₀ and M₂₁ Subunits ofChicken Gizzard PP1_(M).

Polyclonal sheep antibodies to the PP1_(M) holoenzyme were raised in theScottish Antibody Production Unit (Carluke, Ayrshire, U.K.). Antibodieswhich recognise the M₁₁₀ subunit specifically were obtained by passingthe antiserum down a 4 ml affinity column comprising 40 mg ofM₁₁₀-(R714-I1004) coupled covalently to 1 g of dried CNBr-activatedSepharose 4B (Sigma). After washing with 10 column volumes of 50 mMTris/HCl pH 7.5, 1% (by mass) Triton X-100, 0.1 mM EGTA, 0.1% (by vol)2-mercaptoethanol (Buffer B) plus 0.5 M NaCl, followed by 10 volumes ofBuffer B plus 1 M LiBr, the anti-M₁₁₀ antibody was eluted with 50 mMglycine pH 2.0, neutralised immediately with 1 M Tris/HCl pH 8.0 andstored in aliquots at −80° C. Antibodies which recognise the M₂₁ subunitspecifically were obtained in an identical manner, except that theaffinity column comprised about 40 mg of the expressed chicken gizzardM₂₁ subunit coupled to 6 g (dry weight) of CNBr-activated Sepharose.

Removal of the M₂₁ Subunit From PP1_(M).

PP1_(M) (0.01 ml, 0.4U/ml) was dissociated by incubation for 30 minuteswith 500 μM arachidonic acid [25] and then for 30 minutes with 0.08 mlof packed Protein G-Sepharose coupled to 0.08 mg of affinity purifiedanti-M₂₁ antibody. The Protein G-Sepharose was pelleted, and thesupernatant diluted at least 15-fold to allow the M₁₁₀ subunit and PP1cto recombine. The M₁₁₀-PP1c complex was further purified by gelfiltration on Superose 12 (30×1 cm) to ensure complete removal of anyfree PP1c.

Results.

Identification of a PP1c-interaction Domain on the G_(M)-subunit ofPP1_(GM).

The amino acid sequence of rat hepatic G_(L) is 23% identical (39%similar) to residues 1-286 of G_(M) from human skeletal muscle [7].There is no homology over the first 63 residues but identity is >40%over the regions 63-86, 144-166 and 186-227 of G_(M) suggesting that oneor more of these sequences comprise a PP1-binding domain. Fusionproteins in which GST was linked to fragments of G_(M) were thereforetested for their ability to bind to PP1c. GST-G_(M)-(E2-D118) (FIG. 1)and GST-G_(M)-(E2-P243) (data not shown), but not GST-G_(M)-(H100-P350)or GST itself (FIG. 1) interacted with PP1 in Far Western experiments,indicating that the first 118 residues of G_(M) contain a PP1c-bindingdomain. Moreover, a proteolytic fragment derived fromGST-G_(M)-(E2-D118) whose molecular mass was 5 kDa less thanGST-G_(M)-(E2-D118), but not a proteolytic fragment that was 6 kDasmaller, also interacted with PP1c (FIG. 1). Taken together, theobservations suggested that the region comprising residues 63-86 waslikely to bind to PP1c. We therefore synthesised G_(M)-(G63-T93) andexamined its effect on the enzymatic properties of PP1_(GL), the form ofPP1 associated with rat hepatic protein-glycogen particles.

The interaction of PP1c with G_(L) suppresses the dephosphorylation ofmuscle glycogen phosphorylase by 80% and enhances the dephosphorylationof muscle glycogen synthase by 2-3 fold [21, 26]. Disruption of thecharacteristic properties of hepatic PP1_(GL) can therefore be monitoredvery simply by changes in its specificity. G_(M)-(G63-T93) induced asixfold increase in the phosphorylase phosphatase activity of PP1_(GL),the concentrations required for 50% activation being 30 nM (FIG. 2).G_(M)-(G63-T93) also prevented bacterially expressed GST-G_(L) fromsuppressing the phosphorylase phosphatase activity of PP1c (data notshown). However, G_(M)-(G63-T93) had no effect on the glycogen synthasephosphatase activity of PP1_(GL), nor was there any alteration of theother characteristic properties of PP1_(GL), namely allostericinhibition of the glycogen synthase phosphatase activity byphosphorylase a and binding to glycogen (data not shown). Thus theinteraction of G_(M)-(G63-T93) with PP1_(GL) does not displace G_(L)from PP1c.

G_(M)-(G63-T93) also increased the phosphorylase phosphatase activity ofPP1c, indicating that it binds to PP1c, rather than to G_(L). However,the maximal stimulation was only 37+1.4% (SEM for three experiments),establishing that far greater activation of PP1_(GL) is explained by theability of G_(M)-(G63-T93) to overcome the suppressive effect of G_(L)on the phosphorylase phosphatase activity of PP1c. Several otherpeptides, including a 32 residue peptide related to the C-terminus ofribosomal protein S6([G245,G246]S6[218-249]), G_(M)-(S40-Y55) andG_(M)-(E81-T93) (data not shown), had no effect on the phosphorylasephosphatase activity of PP1_(GL) or PP1c at concentrations up to 10 μM.

The peptides G_(M)-(G63-K80) and G_(M)-(G63-N75) also increased thephosphorylase phosphatase activity of PP1_(GL), but were less effectivethan G_(M)-(G63-T93) and higher concentrations were needed (FIG. 2).G_(M)-(G63-K80) and G_(M)-(G63-N75) did not increase the phosphorylasephosphatase activity of PP1c significantly at concentrations up to 10 μM(data not shown). The phosphorylation of G_(M) at Ser67 by cyclicAMP-dependent protein kinase (PKA) triggers the dissociation of PP1 fromG_(M) in vitro and in vivo [18] and phosphorylation of the peptideG_(M)-(G63-N75) at Ser67 prevented it from increasing the phosphorylasephosphatase activity of PP1_(GL) (FIG. 2A). The increase inphosphorylase phosphatase activity observed at the highestphosphopeptide concentrations (10 μM) may be explained by tracecontamination (<10%) with dephosphopeptide, resulting either fromincomplete phosphorylation of Ser67 or slight dephosphorylation duringthe assay.

Identification of a PP1-interaction Domain on the M₁₁₀ Subunit.

Antibodies were prepared that recognised either the M₁₁₀ or M₂₁ subunitsof the myosin-associated form of PP1 (PP1_(M)) from chicken gizzard(FIG. 3A). Removal of the M₂₁ subunit using the M₂₁-specific antibody(FIG. 3B and see Methods) did not affect the activity of PP1_(M) towardsMLC₂₀ or phosphorylase, the MLC₂₀ phosphatase:phosphorylase phosphataseactivity ratio (0.95±0.03) remaining 15-fold higher than PP1c (FIG. 3B).The M₂₁ subunit bound to M₁₁₀, but had no effect on the MLC₂₀phosphatase or phosphorylase phosphatase activity of PP1c and did notbind to PP1c (D. Johnson unpublished). Thus M₁₁₀ is solely responsiblefor enhancing the dephosphorylation of MLC₂₀ and suppressing thedephosphorylation of glycogen phosphorylase by PP1c [9].

In order to identify which region(s) of M₁₁₀, modulates the specificityof PP1c, fusion proteins were constructed consisting of glutathioneS-transferase (GST) followed by fragments of the M₁₁₀ subunit. Afterexpression in E. coli and purification by affinity chromatography onglutathione-Sepharose, the fusion proteins were cleaved with thrombin torelease GST from fragments of the M₁₁₀ subunit (FIG. 4 and see Methods).M₁₁₀-(M1-E309), which contains seven 33 residue ankyrin repeats locatedbetween residues 39-296, modified the specificity of PP1c in a similarmanner to M₁₁₀ itself, increasing activity towards MLC₂₀ about 3-fold(FIG. 5A) and suppressing activity towards glycogen phosphorylase byabout 80% (FIG. 5B). The concentration of M₁₁₀-(M1- E309) required toactivate the MLC₂₀ phosphatase activity maximally (0.1 nM) was similarto the PP1c concentration in the assay, indicating an extremely highaffinity for PP1c. M₁₁₀-(M1-A150) modified the specificity of PP1similarly, but 10-fold higher concentrations were needed compared toM₁₁₀-(M1-E309) (FIGS. 5C and 5D).

If the GST tags were not cleaved with thrombin, a 10-fold higherconcentration of M₁₁₀-(M1-E309) was needed to modulate the substratespecificity of PP1c, while M₁₁₀-(M1-A150) was unable to stimulate theMLC₂₀ phosphatase activity of PP1c at all (data not shown). GST itselfdid not interact with PP1c (FIG. 1), had no effect on either the MLC₂₀phosphatase or phosphorylase phosphatase activity of PP1c (data notshown), and therefore was not removed from the solution after cleavageof the fusion proteins with thrombin.

In contrast to M₁₁₀-(M1-E309), M₁₁₀-(D39-E309) failed to stimulate theMLC₂₀ phosphatase activity of PP1c, or to inhibit its phosphorylasephosphatase activity (FIGS. 5A and 5B), suggesting that the extremeN-terminus of the M₁₁₀ subunit (i.e. before the start of the ankyrinrepeats) might be important in modulating the specificity of PP1c. Thepeptide M₁₁₀-(M1-F38) was therefore synthesized and found to stimulatethe MLC₂₀ phosphatase activity of PP1c to the same extent asM₁₁₀-(M1-E309), although the concentration required for half maximalactivation (10 nM) was at least 100-fold higher (FIG. 5A). M₁₁₀-(M1-F38)stimulated the dephosphorylation of heavy meromyosin in a similar mannerto the dephosphorylation of MLC₂₀ (data not shown). However, likeM₁₁₀-(D39-E309), M₁₁₀-(M1-F38) did not inhibit the phosphorylasephosphatase activity of PP1c (FIG. 5B). These observations suggestedthat residues beyond 38 were needed to suppress phosphorylasephosphatase activity. Consistent with this, M₁₁₀-(L24-Y496) was lesseffective than M₁₁₀-(M1-A150) or M₁₁₀-(M1-E309) in stimulating the MLC₂₀phosphatase activity of PP1c, but inhibited the phosphorylasephosphatase activity of PP1c in a similar manner to M₁₁₀-(M1-A150)(FIGS. 5C and 5D).

Although M₁₁₀-(D39-E309) and M₁₁₀-(M1-F38) bad no effect on thephosphorylase phosphatase activity of PP1c when each peptide wasincluded individually in the assays at concentrations up to 1 μM (FIG.5), a 39±2% inhibition (SEM n=4) was observed when both peptides wereboth present at 1 μM. Surprisingly, M₁₁₀-(D39-E309) prevented (IC50=0.1M) M₁₁₀-(M1-F38) from stimulating the MLC20 phosphatase activity of PP1c(data not shown). Thus M₁₁₀-(D39-E309) plus M₁₁₀-(M1-F38) do notfaithfully mimic the effect of M₁₁₀-(M1-E309).

We have reported previously that the M₁₁₀/M₂₁ complex suppresses thedephosphorylation of glycogen synthase by PP1c [9] and, consistent withthis finding, the dephosphorylation of glycogen synthase was alsoinhibited by M₁₁₀-(M1-E309) (FIG. 6B). However, the dephosphorylation ofglycogen synthase was greatly enhanced by M₁₁₀-(M1-F38) (FIG. 6A).

The Binding of G_(M) and the M₁₁₀ Subunit to PP1c is Mutually Exclusive.

In order to investigate whether G_(M) binds to the same region of PP1cas M₁₁₀, we next examined the effect of G_(M)-(G63-T93) on theproperties of PP1_(M). G_(M)-(G63-T93) at 10 μM increased thephosphorylase phosphatase activity of PP1_(M) by about 7-fold andsuppressed its MLC₂₀ phosphatase activity by 60-65% (FIG. 7A),indicating that the distinctive properties of PP1_(M) had beendisrupted. Gel-filtration experiments confirmed that G^(M)-(G63-T93) haddisplaced the M₁₁₀ subunit from PP1_(M), dissociating it to PP1c (FIGS.7B and 7C). G_(M)-(G63-T93) also prevented M₁₁₀-(M1-F38) orM₁₁₀-(M1-E309) from stimulating the MLC₂₀ phosphatase activity of PP1c(FIG. 8A), and prevented M₁₁₀-(M1-E309) from suppressing thephosphorylase phosphatase activity of PP1c (FIG. 8B).

Conversely, the presence of 10 μM M₁₁₀-(M1-F38) increased thephosphorylase phosphatase activity of PP1_(GL) by 3.5-fold. Thisresulted from the partial dissociation to PP1c, because the enhancedphosphorylase phosphatase activity was not associated with glycogen, butrecovered in the supernatant after centrifugation of theglycogen-protein particles (not shown).

Discussion.

We have identified a region on G_(M) that binds to PP1c (FIG. 9). Thepeptides G_(M)-(G63-T93), G_(M)-(G63-K80) and G_(M)-(G63-N75) allprevented G_(L) from suppressing the dephosphorylation of glycogenphosphorylase by PP1c and two lines of evidence indicate that thesepeptides interact with PP1c and not with G_(L).

Firstly, the PP1c-catalysed dephosphorylation of glycogen phosphorylaseis stimulated slightly by G_(M)-(G63-T93).

Secondly, PP1c crystallises in the presence of G_(M)-(G63-K80) orG_(M)-(G63-N75) in a different form than is observed in the absence ofthese peptides. PKA phosphorylates G_(M) at Ser67 and the introductionof a negative charge directly into the PP1c-binding domain explains whyphosphorylation of Ser67 triggers the dissociation of G_(M) from PP1c[18]. Phosphorylation of G_(M)-(G63-N75) at Ser67 also prevented thispeptide from interacting with PP1 in the PP1_(GL) complex (FIG. 2).

Although G_(M)-(G⁶³-T⁹³) prevented G_(L) from suppressing thedephosphorylation of glycogen phosphorylase by PP1c, it did notdissociate G_(L) from PP1c, nor did it affect the other characteristicproperties of PP1_(GL). Moreover, unlike G_(L), G_(M)-(G63-T93) did notitself suppress the phosphorylase phosphatase activity of PP1c, butactually enhanced it slightly. These observations demonstrate thatanother region(s) on G_(L) must interact with PP1c and that this otherregion(s) may play an important role in modulating the substratespecificity of PP1c. The presence of a second PP1c binding site inG_(M)/G_(L) would be somewhat analogous to the situation found ininhibitor-1 and DARPP which also contain two PP1-binding sites, high(nM) affinity binding being generated by the conjugation of two lowaffinity binding sites that, individually, only interact with PP1 at tMconcentrations [28]. The second PP1c-binding site on G_(M)/G_(L) mightcorrespond to one of the other regions where G_(M) and G_(L) show >40%identity (residues 144-166 and 186-227 of human G_(M)). AlthoughG_(M)-(H100-P350) was not recognised by PP1c in Far Western experiments(FIG. 1) this result is not definitive because G_(M)-(H100-P350) mayonly interact with PP1c weakly. Alternatively, G_(M)-(H100-P350) mightnot fold correctly or fail to renature after SDS/polyacrylamide gelelectrophoresis.

However, it is also possible that residues 144-166 and 186-227 of G_(M)do not represent part of the second PP1c-binding domain, but part of theglycogen-binding domain. In this connection it should be recalled thatresidues 144-166 and 186-227 are the regions showing greatest similarity(25% identity) to GAC1, which appears to be a homologue of G_(M)/G_(L)in budding yeast [7, 27, 28]. Curiously, GAC1 does not contain a regionhomologous to residues 63-93 of G_(M)/G_(L). It would clearly be ofinterest to compare the effect of GAC1 on the enzymatic properties ofPP1c with those of G_(M) and G_(L).

We have also identified a region on the M₁₁₀ subunit that binds to PP1c.An N-terminal fragment, M100-(M1-E309), enhanced the PP1c-catalyseddephosphorylation of MLC₂₀ and suppressed the dephosphorylation ofglycogen phosphorylase in a similar manner to M₁₁₀ itself (FIG. 5).However, unlike M₁₁₀, this fragment does not bind to myosin. Thus theregion which enhances the dephosphorylation of MLC₂₀ is distinct fromthe myosin-binding domain.

The fragment M₁₁₀-(M1-E309) contains seven ankyrin repeats lying betweenresidues 39 and 296. However, M₁₁₀-(D39-E309) was ineffective as anactivator of the MLC₂₀ phosphatase activity of PP1c or as an inhibitorof the phosphorylase phosphatase activity, and this led to the findingthat a peptide comprising the N-terminal 38 residues of the M₁₁₀ subunitenhances the dephosphorylation of MLC₂₀ to the same extent asM₁₁₀-(M1-E309), although with lower potency. However, M₁₁₀-(M1-F38) didnot inhibit the dephosphorylation of glycogen phosphorylase by PP1csuggesting that residues beyond 38 are required to suppress thisactivity. This view was reinforced by the finding that, although neitherM₁₁₀-(M1-F38) nor M₁₁₀-(D39-E309) inhibited the phosphorylasephosphatase activity of PP1c when present individually, inhibition wasobserved in the presence of both peptides. Moreover M₁₁₀-(D39-E309)actually prevented M₁₁₀-(M1-F38) from stimulating the dephosphorylationof MLC₂₀.

These observations suggest that M₁₁₀-(D39-E309) can bind toM₁₁₀-(M1-F38) and/or PP1c. An interaction with PP1c seems likely becauseit has been found that M₁₁₀-(D39-E309) can enhance the phosphorylaseactivity of PP1_(GL). The presence of a second PP1-binding site in theankyrin-repeat domain of the M₁₁₀ subunit is also supported by theobservation that higher concentrations of M₁₁₀-(M1-A150) andM₁₁₀-(M1-E309) are needed to inhibit the phosphorylase phosphataseactivity of PP1c than are required to stimulate its MLC₂₀ phosphataseactivity (see FIG. 5). The presence of at least two PP1-binding sitesmay explain why the M₁₁₀ subunit and PP1c interact at picomolarconcentrations. The ankyrin repeat domain might suppress thedephosphorylation of some substrates (such as glycogen phosphorylase) bya steric mechanism, preventing them from gaining easy access to thecatalytic centre. This scenario could explain why the dephosphorylationof glycogen synthase is greatly enhanced by M₁₁₀-(M1-F38) yet suppressedby M₁₁₀-(M1-E309) (FIG. 6).

G_(M)-(G63-T93) abolished the distinctive properties of PP1_(M) (FIG.7A), prevented M₁₁₀-(M1-F38) or M₁₁₀-(M1-E309) from modulating thesubstrate specificity of PP1c (FIG. 8) and displaced the M₁₁₀ subunitfrom PP1_(M) (FIG. 7B). In addition, the peptide M₁₁₀-(M1-F38), wascapable of displacing G_(L) from PP1_(GL). These findings indicate thatthe binding site(s) on PP1c for G_(M) and the M₁₁₀ subunit are likely tooverlap, explaining why different forms of PP1 contain a singlePP1-targeting subunit. The three-dimensional structure of PP1c isoformshave recently been solved to high resolution [29,30], and PP1ccrystallises in different forms in the presence of G_(M)-(G63-N75) orG_(M)-(G63-K80) or M₁₁₀-(M1-F38) than in the absence of these peptides.

Consistent with the results presented here, Gailly et al [31] haverecently shown that M₁₁₀-(M1-F38) or M₁₁₀(M1-E309) enhance the abilityof PP1c to stimulate the relaxation of microcystin-contractedpermeabilised portal vein, while G_(M)-(G63-T93) inhibits the ability ofPP1_(M) to induce the relaxation of this smooth muscle. G_(M)-(G63-T93)also slowed the relaxation of permeabilised femoral artery, indicatingthat it competes with the endogenous M₁₁₀ subunit for PP1c [31]. Thusthe PP1c-binding peptides described constitute useful pharmacologicalagents with which to explore the role and regulate the activity of PP1in cell regulation.

EXAMPLE 2 Structural Basis for the Recognition of Regulatory Subunits bythe Catalytic Subunit of Protein Phosphatase 1

Materials and Methods

Crystallisation and Data Collection

The catalytic subunit of PP1 1 was overproduced in Escherischia coli andpurified as described previously (Alessi et al., 1993; Barford andKeller, 1994). The G_(M)[G63-N75] peptide, variants of this peptide inwhich Val 66′ or Phe 68′ were changed to Phe, and the peptidesM₁₁₀[1-38] and M₁₁₀[1-35] were synthesised on an Applied Biosystems 430Apeptide synthesiser and purified by chromatography on a C18 column(Johnson et al., 1996) by Mr F. B. Caudwell at Dundee. A three-foldmolar excess of G_(M)[G63-N75] was added to the protein solution (8mg/ml), which had been previously dialysed against 10 mM Tris-HC1 (pH7.8), 0.3 M NaCl, 0.4 mM MnCl₂ and 2 mM DTT. The complex wascrystallised at 20° C. using the hanging drop vapour diffusion method,by mixing 2 ml of the protein-peptide solution and 2 ml of theprecipitant solution containing 2.0 M ammonium sulphate, 2% (w/v)polyethylene glycol 400,100 mM HEPES (pH 7.5) and 2 mM DTT. Theseconditions are very much in contrast to the relatively low ionicstrength conditions from which the monoclinic PP1c crystals grew(Barford and Keller, 1994; Egloff et al., 1995). Crystals appeared after3 months as a cluster. Individual crystals removed from the cluster haddimensions of ˜25 μm×25 μm×5 μm. Crystals were frozen in a 100 Knitrogen gas stream and stored. Prior to freezing, crystals wereincubated in a cryoprotectant solution consisting of an equilibrationbuffer; 2.0 M ammonium sulphate, 2% (w/v) PEG 400, 100 mM HEPES (pH 7.5)with increasing amounts of glycerol in steps of 7%, 15%, 22% and 30%(v/v).

A partial data set to 3.0 Å was collected on Beam Line PX 9.6, SRS,Daresbury, using a 30 cm diameter Mar Research image plate system. Datawere processed and scaled using DENZO and SCALEPACK (Otwinowski, 1993).The crystal system is tetragonal with point group symmetry P422 and unitcell dimensions a=b=62.50 Å, c=361.30 Å. Systematic absences indicate a21 screw axis along b. The Matthews coefficient was 2.38 Å₃ per Dalton,assuming 2 molecules per asymmetric unit. A second data-set wascollected on BL4 at the ESRF, Grenoble. Substantial radiation damage wasobserved during data collection requiring that three crystals were usedin total. Data collected from four crystal at Daresbury and the ESRFwere merged together in SCALEPACK. Details of the data collection andprocessing statistics are given in Table 1.

Structure Determination

The structure of the PP1-G_(M)[63-75] complex was solved by molecularreplacement using as a model the protein atoms coordinates of the 2.5 Årefined structure of the catalytic subunit of PP1γ1 determined by MADmethods (Egloff et al., 1995). Rotation and translation functionssearches were performed with AMORE (Navaza, 1992). Using data between 8and 3 Å resolution, the peak in the rotation search was 6.7 standarddeviations (SD) above the mean. The translation search was bestperformed using data between 8 and 3.5 Å, giving a maximal peak at 13.8SD above the mean for the space group P41212. After the first rigid bodyrefinement performed in AMORE, the R-factor was 0.494 and thecorrelation factor 0.30.

Crystallographic Refinement

The solution from molecular replacement was optimized by 20 cycles ofrigid body refinement performed with X-PLOR version 3.1 (Brunger, 1992),using data between 8.0 Å and 3.0 Å resolution. After a round ofconjugate gradient positional refinement and simulated annealingmolecular dynamics to 2000 K, followed by 25 cycles of grouped B-factorrefinement (2 B-factor groups for each residue), the R factor(respectively free-R) was 0.295 (0.367). Fourier difference maps (Fo-Pc)and (3Fo-2Fc) revealed the presence of three strong peaks at (overthree-times the sigma level of the map) at the catalytic site of PP1c.From the previously refined PP1c-structure, we identified two asmanganese and iron ions. The third one, occupying the position of thetungstate ion in the PP1c-WO4 complex, was identified as sulphate. Theinitial difference Fourier maps also revealed strong electron densitynear the N-terminus of β14. The maps were improved by applyingnon-crystallographic symmetry 2-fold averaging using PHASES (Furey andSwaminathan, 1990). As shown in FIG. 1A, residues Val 66′, Ser 67′ andPhe 68′ of the G_(M)[63-75] peptide were identified in the averaged map.These 3 residues, as well as the 2 metal and sulphate ions were built ineach molecule, using the program TURBO-FRODO (Roussel and Cambillau,1992). Refinement of this structure was performed by repeated rounds ofmanual rebuilding followed by conjugate gradient positional refinementand grouped B-factor refinement using X-PLOR. The final model containsprotein residues Lys 6 to Ala 299 and peptide residues Arg 65′ to Ala69′ in molecule 1, and protein residues Asn 8 to Lys 297 and peptideresidues Gly 63′ to Ala 69′ in molecule 2. A few well defined watermolecules were also observed in both initial (3Fo-2Fc) and (Fo-Fc)electron density maps. Eventually, 14 water molecules that were above 3sigma in the (Fo-Fc) difference map, within hydrogen bond of thePP1-peptide complex or another solvent molecule and present in bothmolecules, were included in the model. The crystallographic andrefinement data are summarized in Table 1. Representative electrondensity from the peptide before and after refinement is shown in FIGS.10A and 10B, respectively. Solvent accessible surface areas werecalculated using the method of Lee and Richards (1974).

Purification and Assay of PP1.

PP1c was isolated from the rabbit skeletal muscle PP1-G_(M) complex asdescribed previously (Johnson et al, 1996). Glycogen particles isolatedfrom rat liver (Schelling el al, 1988) served as the source ofPP1-G_(L). The dephosphorylation of glycogen phosphorylase (10 μM) andthe isolated MLC₂₀ of smooth muscle myosin (1 μM) by PP1c was carriedout as described previously (Cohen et al., 1988; Alessi et al., 1992).

TABLE 1 Crystallographic data and refinement statistics Crystallographicdata: Space group P4₁2₁2 Unit cell parameters (Å) a = b = 62.50; c =361.30 Number of molecules per 2 asymmetric unit Temperature (K) 100Total measured reflections 290671 Number of unique reflections 15509Mean I/s(I) 7.5 Completeness (%) 87 Overall R-merge (%) 14.7 Refinementstatistics: Number of reflections used for 13078 refinement Resolutionrange (Å) 8.0-3.0 R-work 0.223 R-free 0.308 Number of residuesprotein  peptide Molecule 1 294 (Lys 6 to 6 (ARVSFA) (SEQ ID No 21) Ala299) 6 (RRVSFA) SEQ ID No 3) Molecule 2 290 (Asn 8 to Lys 297) R.m.s.d.from ideal bond lengths 0.012 (Å) R.m.s.d. from ideal angles (°) 1.863Number of water molecules Molecule 1 7 Molecule 2 7

TABLE 2 PP1-peptide polar interactions Peptide Protein Water atom atommolecule Distance (Å) Molecule 1 Arg 65' 0 — 7 W 3.2 Val 66' N Asp242OD2 3.0 (**) Ser 67' N Leu 289 O 3.3 Ser 67' OG 7 W 2.7 Ser 67' O Cys291 N (*) 3.2 Ala 69' N Cys 291 O (*) 2.8 Molecule 2 Arg 64' Glu 287 O2.6 NH1 (**) Arg 65' O 7 W 2.8 Val 66' N Asp 242 OD2 3.2 (**) Ser 67' NLeu 289 O (*) 3.1 Ser 67' OG 7 W 2.6 Ser 67' Cys 291 N (*) 3.0 Ala 69' NCys 291 O (*) 3.3 PP1-peptide hydrophobic interactions Peptide Proteinresidues residues Val 66' Ile 169 (*), Leu 243 (*), D242 (**), Leu 289(*), Cys 291 (*) Phe 68' Phe 257 (*), Cys 291 (*), Phe 293 (*) Ala 69'Met290 (**) The star (*) indicates residues absolutely conserved in allprotein phosphatase 1 sequences available so far, the double start (**)the residues mostly conserved (from sequence alignment from Barton etal, 1994).Results and Discussion

Structure Determination.

Crystallographic data to 3.0 Å were measured at the ESRF beam-line BL4at Grenoble and at PX9.6, Daresbury (Table 1). The relatively highmerging R-factors and low I/(I values of the crystallographic dataresults from the weak diffraction observed from the PP1-G_(M)[63-73]crystals. This is attributable to both the small crystal size (˜25 μm by25 μm by 5 μm) and long c-axis of the unit cell. In addition, the highx-ray photon dose required to obtain usable diffraction images resultedin x-ray radiation damage to the crystals, despite being maintained at atemperature of 100 K during the course of the experiment. The structurewas solve by the molecular replacement method using as a search modelthe 2.5 Å refined coordinates of PP1c (Egloff et al., 1995). Phasesobtained from a single cycle of simulated annealing refinement of theprotein coordinates alone using X-PLOR Brunger, 1992), and improved by2-fold non-crystallographic symmetry averaging and solvent flattening,were used to calculate an electron density map. This map revealed cleardensity corresponding to residues Val 66′, Ser 67′ and Phe 68′ (where ′denotes residues of the peptide) of the G_(M) peptide and provided astarting point for further refinement of the PP1-G_(M) peptide complex(FIG. 10A). The final model of the complex was refined at 3.0 Åresolution with a crystallographic R-factor of 0.22 and R-free of 0.31(FIG. 10B). The two molecules of PP1c within the asymmetric unit aresimilar with a root mean square deviation between main chain atoms of0.6 Å. Residues 6 to 299 and 8 to 297 from molecules 1 and 2respectively, are visible in the electron density map. Similar to thestructures of native PP_(γ1) (Egloff et al., 1995) and PP1α in complexwith microcystin LR (Goldberg et al., 1995), residues C-terminal to 299are disordered.

Overall Structure of PP1

The conformation of PP1c in the PP1-G_(M) complex is virtually identicalto that of native PP1c in complex with tungstate (Egloff et al., 1995)with a root mean square deviation between equivalent main-chain atoms of1.0 Å. PP1c is folded into a single elliptical domain consisting of acentral β-sandwich of two mixed β-sheets surrounded on one side by7α-helices and on the other by a sub-domain consisting of 3α-helices anda 3 stranded mixed α-sheet (FIG. 2A, B). The interface of the threeβ-sheets at the top of the β-sandwich creates a shallow catalytic sitechannel. Three loops connecting β-strands with α-helices within aβ-α-β-α-β motif in sheet 1 (strand order β4-β3-β2-β13-β14) together withloops emanating from the opposite β-sheet (sheet 2; strand order,β1-β5-β6-β10-β12-β11) provide the catalytic site residues. The catalyticsite of PP1 contains a binuclear metal site consisting of Mn²⁺ and Fe²⁺(Egloff et al., 1995) and, in the PP1-G_(M) complex, oxygen atoms of asulphate ion of crystallisation coordinate both metal ions, similar tothat seen in the PP1-tungstate (Egloff et al., 1995) and PP2B-phosphatecomplexes (Griffith et al, 1995).

PP1c-G_(M)[63-75] Peptide Interactions

Six residues of the G_(M)[63-75] peptide (Arg 64′ to Ala 69′) areclearly visible in the electron density map of the complex of molecule2, the remaining residues are not visible and assumed to be disordered(FIG. 10B). Density is not visible for Arg 64′ of the peptide bound tomolecule 1, otherwise equivalent residues of the peptide are similarwithin the two complexes. The six residues (RRVSFA) (SEQ ID No 3) of theG_(M)[63-75] peptide in complex 2 adopt an extended conformation andbind to a hydrophobic channel on the protein surface with dimensions 25Å by 10 Å that is formed at the interface of the two β-sheets of theβ-sandwich opposite to the catalytic site channel and therefore remotefrom the catalytic site (FIG. 11A). The residues that form this channeloccur on three regions of PP1c, namely (i) the N-terminus of 5 and β5/β6loop of sheet 2; (ii) the three edge β-strands of sheet 2: β10, β12, β11and (iii) β13, the β13/β14 loop and β14 of the edge of sheet 1 (FIG.11A). The total solvent accessible surface area buried on formation ofthe complex is 980 Å2. Three residues of the peptide (Ser 67′ to Ala69′) form a β-strand which is incorporated into β-sheet 1 of PP1c as asixth β-strand parallel to the N-terminus of the edge β-strand, β14(residues Leu 289 to Leu 296) (FIG. 11C). Main-chain atoms of Ser 67′and Ala 69′ form H-bonds to the main-chain atoms of residues of β14. Inaddition, the main-chain nitrogen of Val 66′ forms a H-bond with theside-chain of Asp 242. Other polar interactions include the guanidiniumgroup of Arg 64′ with the mainchain carbonyl of Glu 287 and a saltbridge to Asp 166. Both Asp 166 and Asp 242 are invariant in mammalianPP1 genes. A water molecule bridges the main-chain carbonyl of Arg 65′and side-chain hydroxyl of Ser 67′ with the main-chain carbonyl of Thr288 of PP1c (FIG. 11C). A notable feature of the peptide binding site isthe presence of a negatively charged region created by seven acidicresidues (with one Lys residue) surrounding the hydrophobic channel atthe N-terminus of the peptide in the vicinity of Arg 64′ and Arg 65′that includes Asp 166 and Asp 242. This would suggest a favourableelectrostatic environment for the side chains of Arg 64′ and Arg 65′.

The predominant interactions between the peptide and PP1c involvehydrophobic contacts between the side chains of Val 66′ and Phe 68′ andsolvent exposed, invariant, hydrophobic residues of PP1c that form thehydrophobic channel (FIG. 11C, E). In particular, the binding site forthe side chain of Val 66′ is formed from the side chains of Ile 169, Leu243, Leu 289 and Cys 291, whereas that for the side chain of Phe 68′ isformed from the side chains of Phe 257, Cys 291 and Phe 293. Details ofpeptide-PP1c contacts are given in Table 2. The structure of theG_(M)[63-75] peptide binding site is likely to be conserved in otherforms of PP1 from diverse species. Each hydrophobic residue of PP1c thatinteracts with the Val 66′ and Phe 68′ residues of the G_(M)[63-75]peptide are invariant and the acidic residues that surround theN-terminus of the peptide binding site are highly conserved amongst allisoforms of PP1 from species as diverse as yeast, Drosophila, mammalsand higher plants (Barton et al., 1994). However, since these residuesare not conserved within the PP2A and PP2B sequences, these proteinswill not recognise PP1-regulatory subunits.

Presence of an (R/K) (V/I)×F Motif in Other PP1c Regulatory Proteins

Over a dozen regulatory subunits of PP1c are now known which appear tobind to PP1c in a mutually exclusive manner that suggests either anoverlapping binding site or sites. Sequence comparisons of thesesubunits reveals little similarity except for the motif (R/K) (V/I)×F,that is not only present in G_(M)[63×75] but also in G_(M), G_(L), M₁₁₀,NIPP-1, p53BP2, and an RNA splicing factor (FIG. 12A). Moreover, a 38residue peptide from the 110kDa M₁₁₀ subunit that binds to PP1c containthis motif (Johnson et al, 1996), as do fragments of NIPP-1 (Beullens etal., 1992; Van Eynde et al, 1995), an RNA splicing factor (Hirano etal., 1996) and p53BP2 (Helps et al., 1995). A 32 residue peptide fromp53BP2, which contains this motif, disrupted the interaction of the M₁₁₀subunit with PP1c, as shown by a decrease in the rate ofdephosphorylation of the MLC₂₀ subunit of smooth muscle myosin and by anincrease in the rate of dephosphorylation of glycogen phosphorylase(FIG. 13A). This peptide also disrupted the interaction of the G_(L)subunit with PP1c, as shown by an increase in the rate ofdephosphorylation of glycogen phosphorylase (FIG. 13B). Peptidescomprising the motif (R/K) (V/I)×F are thus encompassed within the scopeof the invention.

In further support of the notion of a common PP1c recognition motifpresent within PP1-binding proteins, previous studies had revealed thatthe sequence KIQF (SEQ ID No 22) (similar to the R/KVxF motif) at theN-terminus of inhibitor 1 and its homologue DARPP-32 (FIG. 12A) isnecessary for mediating the inhibitory effects of these proteins. Lossof Ile 10 of the KIQF (SEQ ID No 22) motif of inhibitor 1 disrupts theinhibitory effects on PP1c by phospho-inhibitor-1 (Aitken and Cohen,1984; Endo et al., 1996) and the binding of either dephospho-inhibitor-1or phospho-inhibitor-1 to PP1c (Endo et al., 1996). A similar result wasfound on disrupting the equivalent residue (Ile 9) of DARPP-32 (Hemmingset al., 1990; Desdouits et al., 1995). These results were interpreted toindicate that inhibitor-1 and DARPP-32 bind to PP1 through two lowaffinity binding sites, one that encompasses the sequence KIQF (SEQ IDNo 22) and another which includes the phosphorylated Thr residue (35 inI-1, 34 in DARPP-32) and which presumably binds at the catalytic site.Analysis of the PP1-G_(M)[63-75] complex structure suggests that anisoleucine residue could be readily accommodated within the peptidebinding site in place of Val 66′ such that the additional methyl groupon Ile compared to Val would contribute to favourable van der Waalsinteractions between the peptide and Leu 243 and Cys 291 of PP1. Morebulky hydrophobic residues such as Leu, Met and Phe cannot beaccommodated, however. It is interesting to note that as well as the(R/K) (V/I) x F motif shared by PP1-regulatory subunits, the fourresidues N-terminal to this motif contain an abundance of basicresidues. These residues may provide further favourable interactionswith the negative electrostatic surface potential at the N-terminus ofthe G_(M)(63-75) peptide binding site of PP1c.

Mutagenesis of the R/K) (V/I) x F motif

The structural studies presented here suggest a dominant role for Val66′ and Phe 68′ in stabilising the interaction between G_(M)[63-75] andPP1c and this notion is further reinforced by the finding that otherPP1-regulatory subunit sequences contain an (R/K)(V/I) x F motif yetshare little overall sequence similarity. To test the hypothesis thatVal 66′ and Phe 68′ are required for the interaction of G_(M)[63-75]with PP1c and also that the KVKF (SEQ ID No 5) sequence present withinthe M₁₁₀[M1-F38) peptide is important in mediating its interaction withPP1c, we synthesised variations of the G_(M) and M₁₁₀ peptides where theR/KVxF motif was disrupted. The two variants of the G_(M) peptide wereVal 66′ and Phe 68′ to Ala substitutions. In order to disrupt the(R/K)(V/I) x F present within the M₁₁₀ peptide, a peptide correspondingto residues Met 1 to Lys 35 was synthesised which no longer contains thesequence VKF of the VxF motif, which is present at residues 36-38.

The results for the M₁₁₀[1-38] and M₁₁₀[1-35] peptides (FIGS. 14, 15)are unequivocal. Whereas M₁₁₀[1-38] stimulates the myosin light chainphosphatase activity of PP1c with a half-maximal effect at 10 nMreaching maximal (3-fold) activation at a peptide concentration of 1 μMas reported previously (Johnson et al, 1996), the M₁₁₀[1-35] peptide wasat least 104-fold less effective at activating PP1c (FIG. 14). UnlikeM₁₁₀[1-38], the M₁₁₀[1-35] peptide was also unable to activate thephosphorylase phosphatase activity of liver PP1-G_(L). This latterresult suggests two conclusions. Firstly, that although M₁₁₀[1-38] isable to bind to PP1c and disrupt the interactions between PP1c and theG_(L)-subunit, hence reversing the inhibitory effects of G_(L) on theability of PP1c to dephosphorylate phosphorylase, loss of the VKFsequence in the M₁₁₀[1-38] peptide abolishes the ability of the peptideto disrupt this interaction. Secondly, the recognition site on PP1c forthe VKF sequence of the M₁₁₀[1-38] peptide must overlap with the bindingsite for the G_(L) subunit, suggesting that the VKF sequence binds tothe same site as the VSF sequence of G_(L) that is identical with thatpresent in the G_(M)[63-75] peptide. Similar conclusions may be reachedfrom the results obtained from disrupting the VxF motif within theG_(M)[63-75] peptide (FIG. 16B). Substitution of Phe 68′ for Alaabolishes completely the ability of G_(M)[63-75] to disrupt thePP1-G_(L) complex, whereas replacement of Val 66′ with Ala reduced theeffectiveness of the disruption 100-fold.

Thus preferred peptides may comprise analogues of G_(M) withsubstitutions at Val 66′ and Phe 68′ to some other amino acid such asAla, so that binding of the PP1c to G_(M) does not occur and the PP1c isnot suitably directed or controlled. Alternatively, suitable peptidescould comprise peptides suitable to compete for the binding site(s) ofVal 66′ and Phe 68′ on PP1c. Such peptides can be added in sufficientquantities to compete for the Phe 68′ and Val 66′ binding site(s) on thePP1c, thereby disrupting the interaction of PP1c and natural G_(M). Suchpeptides could comprise structural analogues of G_(M) with Phe 68′ andVal 66′ in the same positions as G_(M). Alternatively, other amino acidscapable of mimicking the binding of Phe 68′ and Val 66′ could be used inthese locations.

Regulation of the PP1-G_(M) Complex by Phosphorylation of Ser 67′

Phosphorylation of Ser 67′, corresponding to x of the VxF motif, by PKApromotes dissociiation of both G_(M) and G_(M)[63-75] from PP1c. Invivo, this provides a mechanism of inhibiting PP1c during stimulation ofskeletal muscle by adrenalin (Dent et al., 1990). The sequence of G_(M)surrounding Ser 67′ (RRVSFA) (SEQ ID No 3) conforms to a consensus PKArecognition sequence. Interestingly, the conformation of the peptide issimilar to that of residues 18 to 23 corresponding to thepseudo-substrate sequence of PKI bound to the catalytic site of PKA(Knighton et al., 1990). Although the side chain of Ser 67′ is exposedwithin the PP1c-peptide complex, overall the G_(M) peptide is buried,and it is unlikely that Ser 67′ would be a substrate for PKA when thepeptide is bound to PP1c. This would suggest that PKA phosphorylates Ser67′ when G_(M) is not associated with PP1c and that this phosphorylationprevents the re-association of PP1c with G_(M). Since phosphorylation ofSer 67′ promotes the dissociation of the PP1-G_(M) complex both in vivoand in vitro, it is most likely that PKA phosphorylates Ser 67′ of G_(M)by competing with PP1c for the RRVSFA (SEQ ID No 3) sequence. This isconsistent with a notion that the PP1-G_(M) complex exists in dynamicequilibrium with free PP1c and G_(M) subunits and that phosphorylationoccurs on the regulatory subunit during transient dissociation fromPP1c. In the PP1c-peptide complex, the side-chain of Ser 67′ adopts themost favourable rotamer conformation. Analysis of the PP1c peptidecomplex structure suggests that incorporation of a phosphate group ontothe side chain of Ser 67′ with the same side-chain rotomer conformationwould cause steric hindrance between the peptide and Met 290 of PP1 andalso introduce a phosphate group into a region of negative charge at thePP1c surface (FIG. 11C). This may explain how phosphorylation of Set 67′prevents peptide association with PP1c, although it should be noted thatrotation of the side-chain of Ser 67′ would relieve this steric clash.

A similar mechanism of control may also operate for other PP1-regulatorysubunits. For example, NIPP-1 a nuclear inhibitor of PP1, inhibits PP1with an inhibitory constant of 1 pM (Beullens et al., 1992).Phosphorylation of NIPP-1 by PKA and/or casein kinase 2 in vitroabolishes this inhibition (Beullens et al., 1993; Van Eynde et al.,1994). Although the sites of phosphorylation on NIPP-1 that mediatethese effects are not yet fully characterised it is known that thesesites occur within the central ˜120 residues of NIPP-1 that incorporatesthe (R/K)(V/I) x F motif (Van Eynde et al., 1995). Interestingly, aconsensus phosphorylation site for PKA (RKNS) (SEQ ID No 23) occursimmediately N-terminal to this motif whereas one casein kinase 2consensus phosphorylation site occurs between the Val and Phe of themotif and another occurs immediately C-terminal to the Phe residue(TFSEDDE) (SEQ ID No 24) (Van Eynde et al., 1995) (FIG. 12A). It ispossible that PKA, casein kinase II or other kinases with similarspecificity, release PP1c from inhibition by NIPP-1 by phosphorylatingNIPP-1 at sites that block its interaction with the (R/K)(V/I) x F motifrecognition site on PP1c.

Model of the PP1c-Phospho-Inhibitor 1 Complex

Our model for the interaction of a (R/K)(VII) x F motif with PP1c,together with previous kinetic data suggesting that the sequence KIQF(SEQ ID No 22) of inhibitor-1 (Aitken and Cohen, 1984; Endo et al.,1996) and DARPP-32 (Hemmings et al., 1990; Desdouits et al, 1995)interacts with PP1c, allowed us to construct a plausible model of acomplex of PP1c with phospho-inhibitor 1. The major assumptions of thismodel were (1) the KIQF (SEQ ID No 22) sequence of inhibitor-1 binds tothe same site as RVSF (SEQ ID No 25) of the G_(M)[63-75] sequence and(2) that the phosphothreonine residue 35 of phospho-inhibitor 1 binds atthe phosphate binding site of the PP1c-catalytic site. Secondarystructure predictions of inhibitor 1 (Rost and Sander, 1993; Rost, 1996)suggested that residues 9 to 14 and 23 to 31 adopt β-strand andα-helical conformations, respectively. The prediction of the sequenceKIQF (SEQ ID No 22) as a β-strand is consistent with our assumption thatthis region of inhibitor-1 adopts the same conformation as RVSF (SEQ IDNo 25) of the G_(M) peptide when bound to the VxF recognition site ofPP1c. We have positioned the residues RRPpTP (SEQ ID No 26) encompassingthe pThr 35 site within the catalytic site channel in an extendedconformation, with the phosphate group of the pThr 35 occupying thephosphate binding site of the catalytic site and the Oy-atom of Thr 35equivalent to the solvent exposed oxygen of a dianion that forms aH-bond to the side-chain of the putative general acid His 125 (Egloff etal., 1995; Griffith et al., 1995). The four consecutive Arg residuesN-terminal to pThr 35 interact with Asp and Glu residues within anacidic groove of PP1c formed from the β7/β8 loop on one side and theβ10/β11 loop and β11 strand on the other, similar to that proposed byGoldberg et al., (1995) for their model of DARPP-32 bound to PP1c. Wepropose that residues 20 to 30 of inhibitor-1 form an amphipathic helixwhich folds around the edge of the β-sandwich of PP1c. The N-terminus ofthis helix is disrupted by prolines at residues 19 and 23. Pro 19 andPro 15 are probably responsible for introducing turns into thepolypeptide chain that allows the β-strand encompassing the KIQF (SEQ IDNo 22) sequence (residues 9 to 14) to connect with the α helix. Themodel of the phospho-inhibitor 1-PP1c complex is shown in FIG. 16.

Prediction of PP1 Recognition Motifs in Yeast PP1-Binding Proteins

The residues in mammalian PP1c that interact with the sequence RRVSFA(SEQ ID No 3) are conserved in S. cerevisiae PP1 suggesting that theproteins in S. cerevisiae known to interact with PP1 (reviewed by Stark,1996) probably bind to a similar hydrophobic groove on the surface ofthe enzyme. Examination of their amino acid sequences revealed that anumber of PP1-binding proteins in S. cerevisiae contained putativePP1-binding motifs that were similar to those present in mammalianPP1-binding proteins (FIG. 12A, B). The S. cerevisiae PP1-bindingproteins not only contain a V/I x F motif, but also a basic residueequivalent to Arg 64′ of G_(M)V the residue that contacts Asp 166, Leu289 and the main-chain carbonyl of Glu 287 of PP1c. Several of the S.cerevisiae proteins also contain a further basic residue (His or Lys) atthe position equivalent to Arg 65′ of G_(M). Another striking feature ofthe putative PP1-binding sequences in S. cerevisiae is the presence of abasic amino acid between the Val/Ile and Phe residues, as is also foundin two mammalian PP1-regulatory subunits, the M₁₁₀ subunit and thep53BP2 (FIG. 12A).

The S. cerevisiae proteins GAC1 and PIG2 show some homology to residues140-230 of mammalian G_(M)V and there is genetic and biochemicalevidence that they may function to regulate glycogen metabolism inbudding yeast (Francois et al., 1992). GIP2 also shares sequencesimilarity with residues 140-230 of mammalian G_(M), while YIL045W is anopen reading frame in the S. cerevisiae genome whose predicted aminoacid sequence shows 41% sequence identity to GIP2. YIL045W contains twoputative PP1-binding motifs and site directed mutagenesis will be neededto establish which (if either) of these sequences binds to PP1c. REG1and REG2 are PP1-binding proteins that play a role in cell growth and,in the case of REG1, glucose repression (Tu and Carlson, 1995; Tu etal., 1996; Frederick and Tatchell, 1996). GIP1, which also contains twoputative PP1-binding motifs, is expressed specifically during meiosis,affects the transcription of late meiotic genes and is essential forsporulation (Tu and Carlson, 1996). SCD5 is a PP1-interacting protein(Tu et al., 1996) that was first isolated as a multicopy suppressor ofthe inviability of clathrin heavy chain-deficient yeast (Nelson et al.,1996).

The findings herein demonstrate that the short peptide sequence, the(R/K)(V/I)XF motif, is critical for PP1c to interact with its regulatorysubunits. PP1c (when complexed to its targeting subunits) plays keyroles in the control of many cellular processed and it is reasonable topredict that over 100 pp1-binding proteins may exist in mammalian cells.Protein sequence data-base searching has revealed that the (R/K)(V/I)XFmotifs are found in 10% of proteins. Thus if ˜100 PP1-binding proteinsoccur in mammalian cells, only 1% of proteins with the (R/K)(V/I)XFmotif will be PP1-binding proteins. The reasons why only a few proteinswith the (R/K)(V/I)XF motif bind to PP1 are numerous. For example, notevery residue may be tolerated at position X or immediately N-terminalor C-terminal to this motif. This study has shown that phosphoserine isnot tolerated at position X and it is therefore likely that Asp or Gluwill not be tolerated either. The structure of the PP1-G_(M)[63-75]complex suggests that large hydrophobic residues will also be excludedfrom position X. Moreover, the Val (or Ile) and Phe residues in many(R/K)(V/I)XF motifs will be buried in the hydrophobic core of theprotein and hence be unable to interact with PP1, since this motif ispredicted to form an amphipathic β-strand conformation. Thirdly, many ofthe (R/K)(V/I)XF motifs will be in extracellular proteins orextracelluar domains of transmembranc proteins and hence be unable tobind to PP1. Particular feature so the tertiary structure of PP1-bindingproteins may allow exposure of this motif on the surface to allowinteraction with PP1. Finally, there is evidence that a secondPP1-binding site exists on the G_(M) and M₁₁₀ subunits (Johnson et al.,1996) and the high affinity interaction of PP1c with protein inhibitor-1is generated by the binding of PP1c to two low affinity sites (Desdouitset al., 1995), one of which is the KIQF sequence belonging to the(R/K)(V/I)XF motif.

The question of how regulatory subunits modulate the substratespecificity of PP1c requires the co-crystallisation of PP1c with adiverse array of regulatory subunits and substrates and is beyond thescope of this paper. However, two models to account for this property ofregulatory subunits are that these subunits either alter theconformation of PP1c or simply target PP1 to its substrates. Bothmechanisms may operate in vivo depending on the regulatory subunits andsubstrates. For example, evidence for the former model has recently beenreported for the enhancement of myosin dephosphorylation by a complex ofPP1c and the M₁₁₀ subunit (Johnson et al., 1996, 1997), whereas theenhancement of the dephosphorylation of glycogen phosphorylase andglycogen synthase by the PP1-G_(M) complex is more consistent with thesecond model (Hubbard and Cohen, 1989).

The identification of the (R/K)(V/I)XF motif also suggests a newapproach for determining the physiological roles of PP1-targetingsubunits whose functions arc unknown. Thus mutation of the (R/K)(I/V)XFmotif should disrupt the interaction of many targeting subunits withPP1c without affecting their binding to the target locus. Expression ofthese mutated proteins under an inducible promoter should lead todisplacement of the normal targeting subunit (complexed to PP1c) fromits target locus, without disrupting the functions of any otherPP1c-targeting subunit complex. Finally, the structural informationdescribed here will also facilitate the rational design of drugs thatact by disrupting PP1-targeting subunit interactions.

EXAMPLE 3 Identification of the Regions on the M₁₁₀ Subunit of ProteinPhosphatase 1M that Interact with the M₁₁₀ Subunit and with Myosin

Abbreviations—PP1_(M), myofibril-associated form of protein phosphatase1; PP1c, catalytic subunit of protein phosphatase-1; M₁₁₀ and M₂₁, 110kDa and 21 kDa regulatory subunits of PP1_(M); MBP, maltose-bindingprotein; GST, glutathione-S-transferase.

SUMMARY

We have previously isolated a form of protein phosphatase-1 (PP1_(M))from avian smooth muscle myofibrils which is composed of the catalyticsubunit of PP1 (PP1c) bound to an M-complex consisting of 110 kDa (M₁₁₀)and 21 kDa (M₂₁) subunits. The interaction of PP1c with an N-terminalregion of the M₁₁₀ subunit enhances the dephosphorylation of myosin andsuppresses the dephosphorylation of other substrates [Alessi, D. R.,MacDougall, L. K., Sola, M. M., Ikebe, M. and Cohen, P. (1992) Eur. J.Biochem 210, 1023-1035; Chen, Y. H., Chen, M. X., Alessi, D. R.,Campbell, D. G., Shanahan, C., Cohen, P. and Cohen, P. T. W. (1994) FEBSLett 356, 51-56; Johnson, D. F., Moorhead, G., Caudwell, F. B., Cohen,P., Chen, Y. H., Chen, M. X. and Cohen, P. T. W. (1996) Eur. J. Biochem.239, 317-325]. In this Example we establish that PP1_(M) accounts fornearly all the myosin phosphatase activity in myofibrils, that the M₁₁₀and M₂₁ subunits are present at similar concentrations in themyofibrillar fraction and that these subunits are entirely bound to PP1.We demonstrate that the M₂₁ subunit does not interact with PP1c, butwith the C-terminal 72 residues of the M₁₁₀ subunit, a region which is43% identical to residues 87-161 of the M₂₁ subunit. A fragment of theM₂₁ subunit, M₂₁-(M1-L146), lacking the C-terminal leucine zipper, alsobound to the M₁₁₀ subunit, but two other fragments M₂₁-(M1-E110) andM₂₁-(E110-K186) did not. The M₁₁₀ and M₂₁ subunits were both found to bemyosin-binding proteins. The C-terminal 291 residues of the M₁₁₀subunit, but not the C-terminal 72 residues, bound to myosin, but theN-terminal fragments M₁₁₀-(M1-E309) and M₁₁₀-M1-S477) did not. Thus theregion of the M,on subunit which stimulates the dephosphorylation ofmyosin by PP1c is distinct from the region which targets PP1_(M) tomyosin. Remarkably, each myosin dimer was capable of binding about 20moles of M₂₁ subunit and many of the M₂₁-binding sites were located inthe myosin “rod domain”. The potential significance of this observationis discussed.

Introduction

Protein phosphatase-1 (PP1), one of the major serine/threonine-specificprotein phosphatases in eukaryotic cells, is regulated by targettingsubunits that direct it to particular subcellular loci, modify itssubstrate specificity and confer the ability to be regulated byextracellular signals (reviewed in [1, 2]). A significant proportion ofthe PP1 in vertebrate muscle extracts is associated with myofibrils andhas enhanced activity towards the P-light chain of myosin and reducedactivity towards other substrates, such as glycogen phosphorylase [3,4]. When isolated from avian (chicken gizzard) [4, 5] or mammalian (pigbladder) [6] smooth muscle, this form of PP1 (PP1_(M)) was found to becomposed of three subunits, namely the catalytic subunit of PP1 (PP1c)and two other proteins with molecular masses of 110 kDa and 21 kDa,termed the M₁₁₀ and M₂₁ subunits, respectively [4, 5]. The M₁₁₀ subunitis complexed to both PP1c and the M₂₁ subunit [4], and is the componentwhich modulates the substrate specificity of PP1c because selectiveremoval of the M₂₁ subunit from PP1_(M) does not affect the rate atwhich either myosin or glycogen phosphorylase are dephosphorylated [7].

The M₁₁₀ subunit has been cloned from rat aorta [5], chicken gizzard [8]and rat kidney [9] cDNA libraries. The N-terminus of the M₁₁₀ subunitcontains seven ankyrin repeats (residues 39-296 of the rat aortaprotein), while alternative splicing in rat uterus [5] gives rise to twodifferent C-termini (FIG. 17A), termed Rat1 and Rad2 (SEQ ID Nos 30 and31). The C-terminus of Rat1 is virtually identical to the C-terminus ofthe M₁₁₀ subunit from chicken gizzard (SEQ ID No 29) (FIG. 17A). Thesequence of the M₂₁ subunit from chicken gizzard is structurally relatedto the C-terminal region of the M₁₁₀ subunit, the most strikingsimilarities occurring from residues 76-141 of the M₂₁ subunit andresidues 921-984 of the chicken gizzard M₁₁₀ subunit (54% identity, FIG.17B). However, the C-terminal 53 residues of the M₂₁ subunit fromchicken gizzard are strikingly similar (83% identity) to the C-terminal53 residues of the rat aorta sequence, both terminating in a leucinezipper (FIG. 17B, [5]).

Residues 1-309 of the M₁₁₀ subunit from rat aorta, M₁₁₀-(M1-E309), mimicthe intact M₁₁₀ subunit in stimulating the dephosphorylation of myosinand in suppressing the dephosphorylation of glycogen phosphorylase byPP1c, but a slightly shorter construct M₁₁₀-(D39-E309) (which stillcontains the seven ankyrin repeats) is unable to modulate thespecificity of PP1c [7]. This observation led to the finding that theN-terminal 38 residues, M₁₁₀-(M1-F38), bind to PP1c and enhance thedephosphorylation of myosin. However, M₁₁₀-(M1-F38) does not suppressthe dephosphorylation of glycogen phosphorylase, suggesting that theankyrin repeats either contain a second PP1c-binding site or preventglycogen phosphorylase from binding to the active site of PP1c, perhapsby steric hindrance [7].

A 13 residue peptide G_(M)-(G63-N75) from the subunit (G_(M)) whichtargets PP1c to glycogen and the sarcoplasmic reticulum in striatedmuscle, has been co-crystallised with PP1c and the structure of thecomplex solved to 3 Å resolution [2]. These studies showed that ahexapeptide sequence in G_(M)-(G63-N75) (Arg-Arg-Val-Ser-Phe-Ala) (SEQID No 3) binds to a small hydrophobic groove on the surface of PP1cforming a β-sheet which runs parallel to another β-sheet in PP1c.Moreover, inspection of other mammalian PP1c-binding proteins revealsthat almost all contain an Arg/Lys-Val/Ile-Xaa-Phe motif that is likelyto be critical for interaction with PP1c [2]. For example, aLys-Val-Lys-Phe (SEQ ID No 5) motif is present between residues 35 and38 of the M₁₁₀ subunit and the deletion of residues 36-38 fromM₁₁₀-(M1-F38) prevents this peptide from stimulating thedephosphorylation of myosin, and from disrupting the interaction of PP1cwith other targetting subunits [2].

The finding that a region near the N-terminus of the M₁₁₀ subunit bindsto PP1c and modulates its specificity raised the question of whichregion on the M₁₁₀ subunit interacted with the M₂₁ subunit, and how thePP1_(M) complex is targeted to the myofibrils. In this Example weidentify regions near the C-terminus of the M₁₁₀ subunit that interactwith the M₂₁ subunit and with myosin, and demonstrate that the M₂₁subunit is also a myosin-binding protein. These findings indicate thatthe domain of the M₁₁₀ subunit which enhances the dephosphorylation ofthe myosin P-light chain is distinct from the region which targets PP1cto the contractile apparatus.

Materials and Methods

Materials

PP1_(M) [4] and the dephosphorylated form of myosin [10] were isolatedfrom chicken gizzard (SEQ ID No 29), and the rod-domain and lightmeromyosin were obtained by subdigestion of chicken gizzard myosin withpapain and chymotrypsin, respectively [11]. PP1_(G) was purified fromrabbit skeletal muscle by Dr G. Moorhead in this laboratory [12] andPP1c dissociated from the glycogen-binding subunit by incubation for 2 hin 2 M LiBr and then purified by gel-filtration on a 30×1 cm column ofSuperose 12 (Pharmacia, Milton Keynes, UK) in the presence of LiBr (0.5M). All other chemicals were from BDH Chemicals (Poole, UK) or Sigma(Poole, UK).

Construction of Vectors for the Expression of Fragments of the M₁₁₀Subunit from Rat Aorta (Rat2 Sequence in FIG. 17A) asGlutathione-S-transferase (GST) Fusion Proteins in E. coli.

A construct pGEX-M₁₁₀-(M1-E309) for the expression of GST-M₁₁₀-(M1-E309)from rat aorta was produced as described previously [7]. A construct forthe expression of GST-M₁₁₀-(M1-S477) was prepared by subcloning aXhoI-HindIII fragment (encoding L24-S477) of pKS-M₁₁₀-(M1-S477)described in [5] into the same sites of pGEX-M₁₁₀-(M1-E309). Theresulting construct expressed a GST-M₁₁₀-(M1-S477) fusion protein withthe additional amino acids SAANSISSLIHRD*(SEQ ID No 27) after S477. Anexpression construct for GST-M₁₁₀-(M377-K976) was produced by deleting aNcoI-NcoI fragment of the construct pGEX-M₁₁₀-(L24-K976) [7].

Construction of Vectors for the Expression of C-terminal Fragments ofthe M₁₁₀ Subunit from Chicken Gizzard (Ch1 Sequence in FIG. 17A, [5]) asMaltose Binding Protein (MBP) Fusion Proteins in E. coli.

A pT7.7 vector for the expression of the C-terminal 291 residues of theM₁₁₀ subunit from chicken gizzard, pT7-M₁₁₀-(R714-I1004) was describedpreviously [7]. A construct for the expression of MBP-M₁₁₀-(R714-I1004)was produced by cloning an NdeI-BamHI fragment of pT7-M₁₁₀-l(R714-I1004)into the pMAL-HA vector (New England Biolabs). Removal of aHindIII-HindIII restriction fragment from pMBP-M₁₁₀-(R714-I1004) allowedexpression of MBP-M₁₁₀-(R714-L934) with the sequence GTGRRFTTS (SEQ IDNo 28) added to its C-terminus. Removal of a NdeI-HindIII restrictionfragment from pMBP-M₁₁₀-(R714-I1004), followed by filling in theoverhanging ends and religating them, allowed expression ofMBP-M₁₁₀-(K933-I1004).

Construction of Vectors for the Expression in E. coli. of the M₂₁Subunit from Chicken Gizzard [5], with and Without the C-terminalLeucine Zipper Domain.

A pT7.7 vector for the expression of the entire coding region (M1-K186)of the M₂₁ subunit was described previously [7]. The leucine zippermotif of the M₂₁ subunit was deleted by removing a SacI-BamHIrestriction fragment from pT7.7 M₂₁, filling in the overhanging ends andreligating them. The construct expressed M₂₁-(M1-R144) with an extra Iand L after residue 144. The M₂₁-(M1-R144) protein was insoluble whenexpressed and was purified as described for the expressed M₂₁ subunit[7].

Construction of Vectors for the Expression of the M₂₁ Subunit fromChicken Gizzard [5] and Fragments of the M₂₁ Subunit asGlutathione-S-transferase (GST) Fusion Proteins in E. coli.

A construct expressing GST-M₂₁ was produced by inserting a NdeI-HindIIIfragment of pT7.7 M₂₁ encoding M1-K186 into the same sites of the pGEXvector modified to include an NdeI site. A construct expressingGST-M₂₁-(M1-E110) plus an additional Ala residue at the C-terminus wasconstructed by deleting a XhoI-HindIII fragment of pGEX-M₂₁, filling inthe overhanging ends and religating them. In order to expressGST-M₂₁-(E110-K186), a NdeI-XhoI restriction fragment of pGEX-M₂₁ wasdeleted and the overhanging ends filled in and religated.

Expression of Proteins in E. coli.

This was carried out essentially as described in [7], except that, afterfreezing the cells at −80° C. and thawing, the lysates were not treatedwith DNAase but sonicated for 4 min on ice (ensuring that thetemperature remained below 4° C.) until the suspension was no longerviscous. The soluble GST-fusion proteins and MBP-fusion proteins werepurified from the supernatant by affinity chromatography onglutathione-Sepharose (Sigma) and amylose resin (New England Biolabs),respectively, according to the instructions of the manufacturers. Afterexpression in E. coli M₁₁₀-(R714-I1004) was the major soluble proteinand all experiments with this fragment were performed using thebacterial extracts.

The chicken gizzard M₂₁ subunit was isolated from E. coli extracts asdescribed [7]. M₂₁ subunit lacking the leucine zipper domain,M₂₁-(M1-L146), like the M₂₁ subunit itself, was obtained in inclusionbodies and therefore recovered in the pellet obtained aftercentrifugation of the bacterial lysates for 30 min at 28000×g. Theinclusion bodies were washed three times in 50 mM Tris/HCl pH 7.5, 0.1MNaCl, 10 mM EDTA, 0.1% (by vol) 2-mercaptoethanol, 1 mM benzamidine, 0.2mM phenylmethylsulphonyl fluoride and 0.5% (by mass) Triton X-100, thenresuspended in 50 mM Tris/HCl pH 7.5, 1 mM EDTA, 1 mM EGTA, 0.03% (bymass) Brij-35, 0.1% (by vol) 2-mercaptoethanol. An aliquot (containing 3mg protein) was made 0.5% (by vol) in trifluoroacetic acid, sonicated,centrifuged for 2 min at 13,000×g and the supernatant (containing thesolubilised M₂₁ subunit) loaded on to a Vydac C18 column (SeparationsGroup, Hesperia, Calif., USA) equilibrated in 0.1% (by vol)trifluoroacetic acid. The column was developed with a linearacetonitrile gradient at a flow rate of 1.0 ml/min with an increase inacetonitrile concentration of 1% per min. Homogeneous M₂₁ subunit, whicheluted at 42% acetonitrile, and M₂₁-(M1-L146) which eluted at 40%acetonitrile were dried in a vacuum concentrator redissolved in water,redried and then dissolved in 50 mM Tris/HCl pH 7.5, 0.1 mM EGTA, 0.03%(by mass) Brij-35, 0.1% (by vol) 2-mercaptoethanol.

Removal of GST and MBP Tags from Fusion Proteins.

GST-M₁₁₀-(1-477) was cleaved with thrombin and the proteinase removedusing benzamidine agarose [7]. GST-M₂₁-(E110-K186) (1 mg/ml) was cleavedby incubation for 1 h at 30° C. with 10 μg/ml thrombin, whileGST-M₂₁-(M1-E110) (1 mg/ml) was cleaved by incubation for 3 h at 30° C.with 1 μg/ml thrombin, because it was more susceptible to degradation bythrombin. MBP-M₁₁₀(K933-I1004) (1 mg/ml) was cleaved by incubation for 8h at 23° C. with Factor Xa (10 μg/ml). Other conditions and removal ofFactor Xa were carried out as described for thrombin [7].

Preparation of Phosphorylated Myosin P-light Chain and PhosphataseAssays.

³²P-labelled myosin P-light chains containing 1.0 mol phosphate per molsubunit was prepared by phosphorylation with smooth muscle myosin lightchain kinase [4]. The dephosphorylation of myosin P-light chain (1 μM)was carried out as in [4] and one unit of activity (U) was that amountwhich catalysed the release of 1 μmole of phosphate in one min. Whenassaying PP1_(M) in immunoprecipitates from the myofibrillar extracts,the tubes were shaken continuously and 3 nM okadaic acid was included toinhibit any PP2A present.

Immunoprecipitation of PP1_(M) from Myofibrillar Extracts.

Antibodies raised against the PP1_(M) holoenzyme (1 μg), which recogniseboth the M₁₁₀ and M₂₁ subunits, but not PP1c, affinity purifiedantibodies specific for either the M₁₁₀ subunit or M₂₁ subunit (5 μg)[7], and control IgG (5 μg) were conjugated separately to 10 μl ofpelleted protein G-Sepharose. After incubation for 30 min at 4° C., theProtein G-Sepharose-antibody conjugate was washed three times with 50 mMTris/HCl pH 7.5, 0.1 mM EGTA, 0.03% (by mass) Brij-35, 0.3M NaCl, 0.1%(by vol) 2-mercaptoethanol before addition of a 100 μl of myofibrillarextract (prepared as in [4]) which had been diluted 10-fold in 50 mMTris/HCl pH 7.5, 0.1 mM EGTA, 0.1% (by vol) 2-mercaptoethanol, 0.2 mMphenylmethylsulphonyl fluoride, 1 mM benzamidine, 10 μg/ml leupeptincontaining 1 mg/ml bovine serum albumin. After incubation for 1 h at 4°C., with shaking, a 10 μl aliquot of the suspension was removed tomeasure the total activity. The remaining 90 μl was centrifuged for 1min at 13,000×g, the supernatant was removed, and the pellet washedtwice in dilution buffer containing 0.2 M NaCl and 0.03% (by mass)Brij-35 (but no bovine serum albumin), once in dilution buffer and thenresuspended in 90 μl of dilution buffer. Myosin P-light chainphosphatase activity was then measured in the supernatant and theresuspended pellet at a further 30-fold final dilution.

Myosin binding assays. Myosin (0.5 mg/ml, 1 μM in terms of myosin heavychains) in 10 mM Hepes pH 7.5, 50 mM KCl, 5 mM MgCl2, 0.1% (by vol)2-mercaptoethanol, was mixed with PP1_(M), M₂₁ subunit, or fragments ofthe M₁₁₀ and M₂₁ subunits, at the concentrations indicated in figurelegends. After incubation for 15 min at 0° C., the solutions werecentrifuged for 2 min at 13,000×g, the supernatants removed, and thepellets washed twice in 10 mM Hepes pH 7.5, 50 mM KCl, 5 mM MgCl2, 0.1%(by vol) 2-mercaptoethanol before resuspension in 50 mM Tris-HCl pH 7.5,0.1 mM EGTA, 0.03% (by mass) Brij 35, 0.6 M NaCl, 0.1% (by vol)2-mercaptoethanol. Aliquots of the supernatant, the resuspended pelletand the suspension before centrifugation were either assayed for myosinP-light chain phosphatase activity or denatured in SDS and analysed bySDS/polyacrylamide gel electrophoresis.

Preparation of a Complex Between GST-M₂₁ and M₁₁₀-(R714-I1004).

GST-M₂₁ (10 μg) was mixed with 80 μl of bacterial extract expressingM₁₁₀-(R714-I1004). After incubation for 15 min at ambient temperaturethe solution was added to 20 μl (packed volume) of glutathione-Sepharoseequilibrated in 50 mM Tris HCl pH 7.5, 0.1 mM EGTA, 0.03% (by mass) Brij35, 0.1% (by vol) 2-mercaptoethanol, 0.2 mM phenylmethylsulphonylfluoride, 1 mM benzamidine and 0.15 M NaCl. After incubation for 30 minat 4° C. with shaking, the Sepharose was washed three times in the samebuffer before eluting the complex with buffer containing 20 mMglutathione pH 8.0.

Other Procedures.

Proteins were labelled with digoxigenin and Far Western analyses carriedout as described [4], except that the digoxigenin-labelled probe wasused at a concentration of 0.2 μg/ml instead of 2 μg/ml.SDS/polyacrylamide gel electrophoresis was carried out on 7.5-15% gelsaccording to Laemmli [13] and on 16.5% gels according to Schagger andvon Jagow [14]. Protein was estimated according to Bradford [15].

Results

PP1_(M) Accounts for Nearly all the Myosin Phosphatase Activity inExtracts Prepared from Chicken Gizzard Moyfibrils.

80-90% of the myosin phosphatase activity present in chicken gizzardhomogenates is recovered in the myofibrils [4]. In the present study, weused antibodies that recognise the M₁₁₀ and/or the M₂₁ subunits ofchicken gizzard PP1_(M) [7] to immunoprecipitate the myosin P-lightchain phosphatase activity from the myofibrillar extracts. About 90% ofthe activity was immunoprecipitated by antibodies raised against thePP1_(M) holoenzyme (FIG. 18A) which recognise both the M₁₁₀ and M₂₁subunits in immunoblotting experiments, but not PP1c. Similarly, about80% of the myosin P-light chain phosphatase activity in the myofibrillarextracts was immunoprecipitated by either the anti-M₁₁₀ antibody or bythe anti-M₂₁ antibody (FIG. 18A). Thus, most of the myosin P-light chainphosphatase activity in myofibrillar extracts is catalysed by PP1cpresent as a complex containing both the M₁₁₀ and the M₂₁ subunits.

Immunoblotting experiments demonstrated that the ratio M₁₁₀:M₂₁ inmyofibrillar extracts was identical to the ratio of these subunits inpurified PP1_(M) (FIG. 18B), which is 1:1 [4]. These immunoblottingexperiments also demonstrated that PP1_(M) comprises 0.1% of the proteinin the myofibrillar extract (see legend to FIG. 18B), identical to theproportion estimated from the fold-purification needed to obtain purePP1_(M) from this fraction (see Table 1 in Ref 4). These experimentsimply that PP1_(M) accounts for virtually all the myosin phosphataseactivity associated with myofibrils, and that neither the M₁₁₀ nor theM₂₁ subunit is present in a significant molar excess over PP1c in themyofibrils.

Identification of a Region on the M₁₁₀ Subunit that Binds to the M₂₁Subunit.

PP1_(M) and several fragments of the M₁₁₀ subunit, were subjected toSDS/polyacrylamide gel electrophoresis (FIG. 19A) and, after transfer tonitrocellulose, the blots were probed with digoxigenin-labelled M₂₁subunit (FIG. 19B). These experiments showed that the M₂₁ subunitrecognised the full length M₁₁₀ subunit (FIG. 19B, track 1),M₁₁₀-(R714-I1004) (FIG. 19B, tracks 2 and 3) and M₁₁₀-(K933-I1004) (FIG.19B, track 5), but not M₁₁₀-(R714-L934) (FIG. 19B, track 4),M₁₁₀-(M1-E309) (FIG. 19B, track 7) or M₁₁₀-(M1-S477) (FIG. 19B, track8). Thus, the M₂₁ subunit binds to the C-terminal 72 residues of theM₁₁₀ subunit. The specificity of this interaction was indicated by theobservation that digoxigenin-labelled M₂₁ subunit recognised onlyM₁₁₀-(R714-I1004) and no other protein in the E. coli extract (track 2in FIGS. 19A and 19B), nor did it recognise the MBP or GST tags, PP1c(FIGS. 19A and 19B) or any of the molecular mass markers (data notshown).

Consistent with the results in FIG. 19, digoxigenin-labelledMBP-M₁₁₀-(R714-I1004) (data not shown) and MBP-M₁₁₀-(K933-I1004) (FIG.20B), but not digoxigenin-labelled MBP-M₁₁₀-(R714-L934) (data notshown), recognised the full length M₂₁ subunit and M₂₁(M1-L146) in FarWestern blotting experiments.

The Region of the M₂₁ Subunit that Interacts with the M₁₁₀ Subunit.

Digoxigenin-labelled M₂₁-(M1-L146) recognised the same fragments of theM₁₁₀ subunit as the full length M₂₁ protein (FIG. 19C), demonstratingthat the C-terminal leucine zipper of the M₂₁ subunit is not requiredfor interaction with the M₁₁₁₀ subunit. However, neitherdigoxigenin-labelled GST-M₂₁-(M1-E110) nor digoxigenin-labelledGST-M₂₁-(E110-K186) recognised M₁₁₀-(K933-I1004) in Far Western blottingexperiments (data not shown). Consistent with these findings,digoxigenin-labelled M₁₁₀-(K933-I1004) recognised the full length M₂₁protein and M₂₁-(M1-L146), but not M₂₁-(M1-E110) or M₂₁-(E110-K186) inFar Western blotting experiments (FIGS. 20A and B). However,digoxigenin-labelled M₁₁₀-(K933-I1004) did recognise a proteolyticfragment of the M₂₁ subunit with an apparent molecular mass onlyslightly larger than M₂₁-(M1-E110) (FIG. 20B, track 2 and compare tracks2 and 4 in FIG. 20A). These results are considered further underDiscussion.

The Isolated M₂₁ Subunit Dimerizes and the Region Involved inDimerization is Identical to that which Interacts with the M₁₁₀ Subunit.

Although the M₁₁₀ subunit binds to both PP1c and the M₂₁ subunit [4],and removal of the M₂₁ subunit does not alter the specificity of thePP1_(M) complex [7], an interaction between the M₂₁ subunit and PP1c hadnot been excluded. In order to examine this point, PP1c and the M₂₁subunit were mixed together and subjected to gel filtration on Superose12. The M₂₁ subunit eluted just before the 37 kDa PP1c protein,demonstrating that they do not form a high affinity complex andsuggesting that the isolated M₂₁ subunit dimerizes (data not shown).These results were supported by the finding that digoxigenin-labelledfull length M₂₁ subunit recognised the M₂₁ subunit as well as the M₁₁₀subunit, but not PP1c, in Far Western blotting experiments (FIG. 21,track 1). Similar results were obtained with M₂₁-(M1-L146) (FIG. 21,track 2). Digoxigenin-labelled M₂₁ subunit, like digoxigenin-labelledM₁₁₀-(K933-I1004), recognised a fragment of the M₂₁ subunit thatmigrated slightly more slowly than M₂₁-(M1-E110), but did not recogniseM₂₁-(M1-E110) or M₂₁-(E110-K186) (Tracks 2, 4 and 5 in FIGS. 20B and20C).

Identification of a Region on the M₁₁₀ Subunit that Binds to Myosin.

When PP1_(M) (30 nM) was mixed with chicken gizzard myosin (1 μM) andcentrifuged to pellet the myosin, 85% of the myosin P-light chainphosphatase was recovered in the pellet (FIGS. 22 and 23A). In contrast,neither PP1c (FIG. 22) nor bovine serum albumin (data not shown) boundto myosin under these conditions. After removal of the M₂₁ subunit fromPP1_(M) [7], the M₁₁₀-PP1c complex (PP1_(M)(ΔM₂₁ ) still pelleted withmyosin in a similar manner to PP1_(M) itself (FIG. 22), indicating thatthe M₁₁₀ subunit is a myosin-binding protein.

In order to identify the myosin-binding domain(s), several fragments ofthe M₁₁₀ subunit were expressed and purified from E. coli extracts andtheir binding to myosin was studied. GST-M₁₁₀-(M1-S477), likeGST-M₁₁₀-(M1-E309) [7], stimulated the PP1c-catalysed dephosphorylationof the myosin P-light chain and inhibited the dephosphorylation ofglycogen phosphorylase in a similar manner to the full length M₁₁₀subunit (data not shown). However, neither GST-M₁₁₀-(M1-S477) norGST-M₁₁₀-(M1-E309) bound to myosin (data not shown), even after removalof the GST-tag from GST-M₁₁₀-(M1-S477) (FIG. 23A).

A fragment comprising GST-M₁₁₀-(M377-K976) from rat aorta migrated asmultiple bands on SDS/polyacrylamide gels after purification onglutathione-Sepharose (FIG. 23A), indicating cleavage at multiple siteswithin the M₁₁₀ subunit. Only the largest fragment, with an apparentmolecular mass corresponding to undegraded GST-M₁₁₀-(M377-K976) bound tomyosin (FIG. 23A), suggesting that the myosin binding site(s) waslocated towards the C-terminus of the M₁₁₀ subunit. Consistent with thisfinding, M₁₁₀-(R714-I1004) from chicken gizzard also bound to myosin(FIG. 23B). However, M₁₁₀-(K933-I1004), which bound to the M₂₁ subunit(FIG. 20B), did not bind to myosin in these experiments (FIG. 23B).

The M₂₁ Subunit, and a Complex Between M₂₁ and M₁₁₀-(R714-I1004) Bind toMyosin.

After purification on glutathione-Sepharose, GST-M₂₁ migrated as fourprotein staining bands (track 1 in FIG. 20A), the two species of highestapparent molecular mass being recognised by the anti-M₂₁ antibody (FIG.23B). The apparent molecular mass of the slowest migrating band (47 kDa)corresponds to undegraded GST-M₂₁ and this species bound to myosin (FIG.23B). The next most slowly migrating band had an apparent molecular massof 38 kDa, slightly less than that of GST-M₂₁-(M1-E110) (data not shown)indicating that it corresponds to GST fused to about the first 100residues of the M₂₁ subunit; this fragment hardly bound to myosin (FIG.23B).

Bacterial extracts expressing M₁₁₀-(R714-I1004) were mixed with GST-M₂₁and the resulting complex was purified on glutathione-Sepharose. Thiscomplex bound quantitatively to myosin (FIG. 23B). In contrast, theGST-M₂₁ fragment of apparent molecular mass 38 kDa was not complexed toM₁₁₀-(R714-I1004) and did not bind to myosin (FIG. 23B). The C-terminalfragment of the M₂₁ subunit, M₂₁-(E110-K186) also did not bind to myosinunder these conditions (data not shown).

Multiple Binding Sites for the M₂₁ Subunit on the Myosin Molecule.

The molar ratio myosin:PP1_(M) in chicken gizzard is about 80:1 in vivo[4] and the myosin binding experiments described above were thereforecarried out using a large (ten fold) molar excess of myosin over eitherthe M₂₁ or the M₁₁₀ subunit. However, further experiments carried outwith the M₂₁ subunit in excess revealed that, remarkably, 20 or moremoles of M₂₁ subunit could be bound to each myosin dimer (FIG. 24A).Many of the binding sites were located in the region of myosin involvedin filament formation, because the M₂₁ subunit was pelleted with themyosin “rod” domain even when the molar ratio M₂₁:myosin dimer was 10:1(FIG. 24B). A shorter portion of the rod, termed light meromyosin, alsobound the M₂₁ subunit avidly. However, a fragment of the M₂₁ subunitlacking the first 15 residues from the N-terminus, which was acontaminant in this preparation, did not bind to light meromyosin (FIG.24B), although it bound to the longer myosin rod (FIG. 24B). The M₂₁subunit lacking the C-terminal leucine zipper, M₂₁-(M1-L146), bound toboth myosin and the rod domain, but fewer moles of M₂₁-(M1-L146) couldbe bound and this C-terminally truncated species did not bind to lightmeromyosin under the conditions studied (FIG. 24C).

Multiple Forms of the M₁₁₀ Subunit

Comparison of two different clones encoding the M₁₁₀ subunit fromchicken gizzard revealed a 123 bp (41 amino acid) deletion/insertionafter Asn-511 (FIG. 17, [8]). Since the rat aorta sequence [5] showedconsiderable variation from the chicken sequences in this region,compared to the high degree of sequence similarity throughout most ofthe rest of the molecule (FIG. 17), it seemed probable that forms of therat M₁₁₀ subunit also existed that varied in this middle section of theprotein. PCR of the “variable region” of several rat aorta clones gavefragments of either 608 bp or 776 bp. Direct sequencing of thesefragments showed an in frame insertion of 168bp (56 amino acids) afterSer-552 (FIG. 1); i.e. a slightly different position from the deletionreported for the chicken gizzard M₁₁₀ subunit (FIG. 17). Furthermore, adifferent 62 amino acid deletion/insertion in this section is apparentby comparison of the rat aorta sequences with that of the M₁₁₀ proteinfrom rat kidney (FIG. 1) [9]. While it is likely that most of thesevariations arise by alternative splicing of the mNRNA, Southern blottingof rat genomic DNA revealed the presence of two closely related genes(data not shown).

Discussion

The contraction of smooth muscle is triggered by phosphorylation of theP-light chain of myosin catalysed by myosin light chain kinase. However,the identity of the myosin P-light chain phosphatase remained unclearfor many years. In 1992 we reported that 80-90% of the myosinphosphatase activity in chicken gizzard homogenates was associated withmyofibrils and purified a myosin phosphatase to homogeneity from thisfraction [4]. This enzyme, termed PP1_(M), was found to be composed ofthe β-isoform of PP1c (termed the δ-isoform in [16]) and an “M-complex”consisting of two other subunits [4] whose molecular masses were 21 kDa(M₂₁) [5] and 110 kDa (M₁₁₀) [5, 8], respectively. Further evidence thata form of PP1 was the major myosin phosphatase in smooth muscle wasindicated by the finding that tautomycin (a much more potent inhibitorof PP1 than PP2A [17]) stimulated the contraction of permeabilisedmammalian smooth muscle fibres at much lower concentrations than okadaicacid [18] (a much more potent inhibitor of PP2A than PP1 [19]).

Two further pieces of evidence presented in this Example establish thatPP1_(M)accounts for most, if not all, of the myosin phosphatase activityassociated with chicken gizzard myofibrils, reinforcing the view that itis likely to be the major myosin P-light chain phosphatase in vivo.Firstly, nearly all the myosin P-light chain phosphatase activity wasimmunoprecipitated by antibodies that recognise either the M₁₁₀ or theM₂₁ subunit specifically (FIG. 18A). Secondly, PP1_(M) was found torepresent 0.1% of the protein in the myofibrillar extracts whether itsconcentration was calculated from the increase in specific activityneeded for purification to homogeneity [4] or from immunoblottingexperiments with the anti-M₁₁₀ and anti-M₂₁ antibodies (FIG. 18B). Hadanother enzyme been the major myosin phosphatase in the myofibrillarextracts the enrichment estimated by immunoblotting with anti-M₁₁₀ andanti-M₂₁ antibodies would have been much higher.

The experiments presented in FIG. 18 also demonstrate that the M₁₁₀ andM₂₁ subunits are not present in myofibrillar extracts in a significantmolar excess over PP1c and that all the M₁₁₀ subunits are complexed toM₂₁ subunit and vice versa. The M₂₁ subunit was found to bind to theC-terminal 72 residues of the chicken gizzard M₁₁₀ subunit (FIGS. 19 and25), a region whose amino acid sequence is 43% identical to residues87-161 of the M₂₁ subunit (FIG. 17B). The C-terminal leucine zipper ofthe M₂₁ subunit (residues 145-186) is not required for interaction withthe M₁₁₀ subunit, and the site on the M₂₁ subunit which interacts withthe M₁₁₀ subunit lies within about the N-terminal 120 residues (FIG.20B). Interestingly, the same region is essential for the dimerisationof the M₂₁ subunit (compare FIGS. 20B and 20C), suggesting that theregion(s) involved in interaction is probably located between residues60 and 120 of the M₂₁ subunit and 906-965 of the M₁₁₀ subunit fromchicken gizzard; i.e. the regions with greatest amino acid identitybetween these two proteins (FIG. 17). More digoxigenin-labelled M₂₁subunit bound to the M₁₁₀ subunit than to the M₂₁ subunit in Far Westernblotting experiments (FIG. 21), consistent with the observation thatM₁₁₀/M₂₁ heterodimers form in vivo, but not M₂₁/M₂₁ homodimers. Thefinding that the C-terminus of the M₁₁₀ subunit interacts with the M₂₁subunit explains why preparations of PP1_(M) comprising PP1c complexedto N-terminal fragments of the M₁₁₀ subunit do not contain the M₂₁subunit [8, 20].

PP1_(M) binds to the dephosphorylated form of myosin and our datademonstrate that the M₁₁₀ subunit (FIG. 22) and the M₂₁ subunit (FIG.23B and FIG. 24) are both myosin-binding proteins. The C-terminal 600residues of the M₁₁₀ subunit from rat aorta, M₁₁₀-(M377-K976) (FIG. 23A)and the C-terminal 291 residues of the M₁₁₀ subunit from chickengizzard, M₁₁₀-(R714-I1004), bound to myosin, but the C-terminal 72residues of the M₁₁₀ subunit, M₁₁₀-(K933-I1004), did not (FIG. 23B),indicating that a myosin-binding domain is likely to be situated in theM₁₁₀ subunit just N-terminal to the M₂₁-binding domain (FIG. 25). Incontrast, two N-terminal fragments of the M₁₁₀ subunit M₁₁₀-(M1-S477)(FIG. 23A) and M₁₁₀-(M1-E309) (data not shown) did not bind to myosinunder the conditions studied. Since M₁₁₀-(M1-E309) [7] andM₁₁₀-(M1-S477) (data not shown) stimulate the dephosphorylation ofmyosin and inhibit the dephosphorylation of glycogen phosphorylase byPP1c, and in a similar manner to full length M₁₁₀ subunit, these resultsshow that the region of the M₁₁₀ subunit which stimulates thedephosphorylation of the myosin P-light chain is distinct from thatwhich binds the dephosphorylated form of myosin and thereby targetsPP1_(M) to the contractile apparatus.

Digestion of chicken gizzard PP1_(M) with chymotrypsin cleaves the M₁₁₀subunit to a fragment with an apparent molecular mass of 58 kDa and aform of PP1, termed here PP1_(M)*, can then be isolated bygel-filtration which appears to comprise just the 58 kDa fragment andPP1c in a 1:1 molar ratio [8]. The 58 kDa fragment, like the M₁₁₀subunit, has a blocked N-terminus and seven tryptic peptides isolatedwere located between residues 286 and 467, suggesting that the 58 kDafragment corresponds to the N-terminal portion of the M₁₁₀ subunit [8].PP1_(M)* was reported to bind to myosin, albeit less effectively thanPPIM [8], suggesting the presence of a myosin-binding domain within the58 kDa fragment. This result is in apparent conflict with the presentstudy, because the fragment M₁₁₀-(M1-S477), which also migrates onSDS/polyacrylamide gels with an apparent molecular mass of 58 kDa, didnot bind to dephosphorylated myosin under conditions where 80-90% of thePP1_(M) and M₁₁₀-(R714-I1004) was pelleted with myosin (FIG. 23A). Onepossible explanation for this discrepancy is that PP1_(M)* also containssmall myosin-binding fragments from the C-terminus of the M₁₁₀ subunitwhich still interact with the N-terminal 58 kDa fragment, but are toosmall to be detected by SDS/polyacrylamide gel electrophoresis. In aseparate study heavy meromyosin (50 μg) was found to bind partially to 2mg of M₁₁₀-(1-633) coupled to Affigel 15, at very low ionic strength butnot at 150-200 mM NaCl [21]. The significance of this observation isunclear because of the extremely high concentration of the M₁₁₀-(1-633)used in these experiments. The average intracellular concentration ofPP1_(M) in chicken gizzard is about 1 μM, 100-fold lower than theconcentration of myosin. In the present study, we analysed the bindingof the M₁₁₀ subunit and its subfragments (30-100 nM) to myosin (1 μM)using low concentrations of these proteins to try and ensure that onlyhigh affinity binding sites were identified.

The isolated M₂₁ subunit also bound to myosin and up to 20 moles of M₂₁subunit could be bound to each myosin dimer (FIG. 24). Theseobservations indicate that each myosin molecule contains multiplebinding sites for the M₂₁ subunit, many of which are located within the“rod domain” (FIGS. 24B and 24C). In vivo, the molar ratioPP1_(M):myosin is about 1:80 and yet, during muscle relaxation, all themyosin P-light chains can be dephosphorylated by PP1_(M) within seconds.This implies that PP1_(M) must be highly mobile within the myofibrilsand move extremely rapidly from one myosin molecule to another. The “offrates” for binding of PP1_(M) to myosin must therefore be very fast aswell as the “on rates”. It is tempting to speculate that the presence ofmultiple binding sites on myosin for the M₂₁ subunit (and perhaps forthe M₁₁₀ subunit as well) allows PP1_(M) to “slide” rapidly from onemyosin molecule to another.

EXAMPLE 4 Design of Small Molecules to Modulate the Properties of PP1

Table A is a print-out of the atomic coordinates of the proteinphosphatase-l peptide coordinates as deduced in Example 2. The format isProtein Data Bank. The structure of the protein phosphatase-1 catalyticsubunit (PP1c) in complex with a 13-residue peptide (G_(M) peptide)corresponding to a site of interaction between PP1c and the glycogentargeting subunit provides a frame-work for the rational design of smallmolecules to modulate the functions and properties of PP1 in vivo.Knowledge of the structural nature of the interactions between the G_(M)peptide and PP1c allows the design of inhibitors that mimic theseinteractions. These inhibitors may be designed for increased potency,cell permeability and with improved pharmokinetic properties.

Computer graphics systems may be used to design such inhibitors inconjunction with computer graphics modelling software such as SYBILavailable from: Tripos Inc, 16995 S Hanley Road, St Louis, Miss.63144-2913, USA and LUDI available from: Molecular Simulations Inc, 9685Scranton Road, San Diego, Calif. 92121-3752, USA, and in conjunctionwith the atomic coordinates shown in Table A.

EXAMPLE 5 Effect of Peptide Derived from p53BP2 Binding Site to PP1 inVivo

The function of p53BP2 is ascertained by examining the in vivo effect ofpeptides based on the sequence of the p53BP2 binding site to PP1. Thismay be done by reference to the consensus peptide sequence described inthe previous Examples and by reference to the crystal structure inExample 2. The peptide is introduced into cultured cells usingpenetratin available from Appligene. Other importins may also be used.Alternatively cDNA specifying p53BP2 proteins mutant in the p53BP2binding site to PP1 are transfected in cultured cells. The effect ofthese agents on the cell cycle and apoptosis are assessed by a number ofmethods, for example WAF1 ELISA and Nuclear Matrix Protein ELISA assays(Amersham).

The effect of the p53BP2 peptide is to modulate the interaction betweenPP1 and p53BP2 in vivo and affect cell regulation and apoptosis. Thep53BP2 peptide may also be micro-injected into the cell.

TABLE A ATOM 1 N LYS 6 −10.263 46.372 91.126 1.00 53.07 0 ATOM 3 CA LYS6 −9.182 46.177 90.159 1.00 53.07 0 ATOM 4 CB LYS 6 −9.220 47.277 89.0921.00 45.20 0 ATOM 5 CG LYS 6 −10.284 47.095 88.015 1.00 45.20 0 ATOM 6CD LYS 6 −9.809 46.200 86.868 1.00 45.20 0 ATOM 7 CE LYS 6 −8.832 46.91985.927 1.00 45.20 0 ATOM 8 NZ LYS 6 −7.498 47.216 86.540 1.00 45.20 0ATOM 12 C LYS 6 −7.814 46.179 90.835 1.00 53.07 0 ATOM 13 O LYS 6 −6.85445.624 90.303 1.00 45.20 0 ATOM 14 N LEU 7 −7.746 46.816 92.005 1.0042.33 0 ATOM 16 CA LEU 7 −6.527 46.941 92.800 1.00 43.14 0 ATOM 17 CBLEU 7 −6.840 47.599 94.141 1.00 24.45 0 ATOM 18 CG LEU 7 −5.670 47.78295.106 1.00 18.34 0 ATOM 19 CD1 LEU 7 −4.775 48.881 94.589 1.00 25.09 0ATOM 20 CD2 LEU 7 −6.186 48.121 96.496 1.00 22.21 0 ATOM 21 C LEU 7−5.892 45.594 93.063 1.00 42.81 0 ATOM 22 O LEU 7 −6.497 44.723 93.6751.00 23.55 0 ATOM 23 N ASN 8 −4.656 45.424 92.627 1.00 17.03 0 ATOM 25CA ASN 8 −4.000 44.156 92.846 1.00 11.65 0 ATOM 26 CB ASN 8 −3.20443.744 91.610 1.00 18.54 0 ATOM 27 CG ASN 8 −3.486 42.312 91.193 1.0015.30 0 ATOM 28 OD1 ASN 8 −4.643 41.903 91.068 1.00 14.53 0 ATOM 29 ND2ASN 8 −2.429 41.538 90.993 1.00 7.61 0 ATOM 32 C ASN 8 −3.110 44.20794.079 1.00 11.35 0 ATOM 33 O ASN 8 −1.906 44.515 93.985 1.00 12.23 0ATOM 34 N ILE 9 −3.716 43.900 95.232 1.00 12.56 0 ATOM 36 CA ILE 9−3.018 43.894 96.523 1.00 14.41 0 ATOM 37 CB ILE 9 −3.955 43.439 97.6901.00 2.00 0 ATOM 38 CG2 ILE 9 −3.155 42.828 98.812 1.00 2.00 0 ATOM 39CG1 ILE 9 −4.742 44.634 98.244 1.00 2.00 0 ATOM 40 CD1 ILE 9 −3.86545.837 98.581 1.00 2.00 0 ATOM 41 C ILE 9 −1.796 42.996 96.501 1.0010.07 0 ATOM 42 O ILE 9 −0.759 43.351 97.045 1.00 2.00 0 ATOM 43 N ASP10 −1.916 41.845 95.849 1.00 2.00 0 ATOM 45 CA ASP 10 −0.822 40.88795.782 1.00 2.00 0 ATOM 46 CB ASP 10 −1.336 39.562 95.208 1.00 45.77 0ATOM 47 CG ASP 10 −2.234 38.801 96.191 1.00 46.34 0 ATOM 48 OD1 ASP 10−3.054 39.444 96.879 1.00 50.94 0 ATOM 49 OD2 ASP 10 −2.123 37.55896.281 1.00 55.84 0 ATOM 50 C ASP 10 0.426 41.369 95.036 1.00 2.00 0ATOM 51 O ASP 10 1.540 41.181 95.516 1.00 43.44 0 ATOM 52 N SER 11 0.24541.993 93.874 1.00 2.00 0 ATOM 54 CA SER 11 1.387 42.494 93.112 1.002.00 0 ATOM 55 CB SER 11 0.987 42.834 91.678 1.00 24.54 0 ATOM 56 OG SER11 −0.025 43.819 91.653 1.00 26.40 0 ATOM 58 C SER 11 1.964 43.72793.804 1.00 2.00 0 ATOM 59 O SER 11 3.168 44.010 93.702 1.00 23.86 0ATOM 60 N ILE 12 1.099 44.467 94.493 1.00 19.26 0 ATOM 62 CA ILE 121.536 45.641 95.226 1.00 19.26 0 ATOM 63 CB ILE 12 0.345 46.351 95.8781.00 2.00 0 ATOM 64 CG2 ILE 12 0.831 47.364 96.909 1.00 2.00 0 ATOM 65CG1 ILE 12 −0.499 46.986 94.775 1.00 2.00 0 ATOM 66 CD1 ILE 12 −1.72247.687 95.245 1.00 2.00 0 ATOM 67 C ILE 12 2.501 45.112 96.275 1.0019.26 0 ATOM 68 O ILE 12 3.684 45.445 96.264 1.00 2.00 0 ATOM 69 N ILE13 1.987 44.246 97.141 1.00 2.00 0 ATOM 71 CA ILE 13 2.764 43.605 98.1991.00 2.00 0 ATOM 72 CB ILE 13 1.899 42.504 98.897 1.00 2.00 0 ATOM 73CG2 ILE 13 2.747 41.645 99.810 1.00 2.00 0 ATOM 74 CG1 ILE 13 0.76443.154 99.691 1.00 2.00 0 ATOM 75 CD1 ILE 13 −0.213 42.167 100.331 1.002.00 0 ATOM 76 C ILE 13 4.039 42.960 97.602 1.00 2.00 0 ATOM 77 O ILE 135.142 43.099 98.158 1.00 2.00 0 ATOM 78 N GLN 14 3.864 42.278 96.4621.00 2.00 0 ATOM 80 CA GLN 14 4.937 41.582 95.740 1.00 2.00 0 ATOM 81 CBGLN 14 4.415 41.065 94.391 1.00 21.13 0 ATOM 82 CG GLN 14 5.467 40.47093.454 1.00 32.43 0 ATOM 83 CD GLN 14 5.655 41.284 92.171 1.00 28.89 0ATOM 84 OE1 GLN 14 4.726 41.428 91.363 1.00 30.84 0 ATOM 85 NE2 GLN 146.861 41.818 91.977 1.00 29.15 0 ATOM 88 C GLN 14 6.088 42.519 95.5151.00 2.00 0 ATOM 89 O GLN 14 7.206 42.249 95.934 1.00 24.80 0 ATOM 90 NARG 15 5.789 43.625 94.848 1.00 15.16 0 ATOM 92 CA ARG 15 6.776 44.63894.552 1.00 15.16 0 ATOM 93 CB ARG 15 6.122 45.812 93.846 1.00 8.83 0ATOM 94 CG ARG 15 6.530 45.981 92.388 1.00 8.83 0 ATOM 95 CD ARG 155.543 46.882 91.684 1.00 8.83 0 ATOM 96 NE ARG 15 4.195 46.322 91.7611.00 8.83 0 ATOM 98 CZ ARG 15 3.094 46.998 91.465 1.00 8.83 0 ATOM 99NH1 ARG 15 3.178 48.261 91.073 1.00 9.33 0 ATOM 102 NH2 ARG 15 1.90746.413 91.567 1.00 8.83 0 ATOM 105 C ARG 15 7.405 45.106 95.841 1.0015.16 0 ATOM 106 O ARG 15 8.622 45.124 95.952 1.00 13.09 0 ATOM 107 NLEU 16 6.575 45.462 96.820 1.00 2.00 0 ATOM 109 CA LEU 16 7.049 45.92498.124 1.00 2.00 0 ATOM 110 CB LEU 16 5.853 46.215 99.033 1.00 2.00 0ATOM 111 CG LEU 16 4.982 47.420 98.662 1.00 2.00 0 ATOM 112 CD1 LEU 163.630 47.302 99.324 1.00 2.00 0 ATOM 113 CD2 LEU 16 5.664 48.707 99.0561.00 2.00 0 ATOM 114 C LEU 16 8.014 44.942 98.809 1.00 2.00 0 ATOM 115 OLEU 16 9.031 45.354 99.361 1.00 2.00 0 ATOM 116 N LEU 17 7.712 43.65098.770 1.00 12.70 0 ATOM 118 CA LEU 17 8.590 42.652 99.395 1.00 4.33 0ATOM 119 CB LEU 17 7.812 41.387 99.780 1.00 4.50 0 ATOM 120 CG LEU 176.740 41.515 100.838 1.00 4.52 0 ATOM 121 CD1 LEU 17 6.338 40.159101.302 1.00 11.14 0 ATOM 122 CD2 LEU 17 7.285 42.318 101.997 1.00 6.930 ATOM 123 C LEU 17 9.796 42.225 98.548 1.00 7.39 0 ATVH 124 O LEU 1710.751 41.652 99.086 1.00 15.30 0 ATOM 125 N GLU 18 9.758 42.492 97.2381.00 65.13 0 ATOM 127 CA GLU 18 10.847 42.104 96.329 1.00 70.48 0 ATOM128 CD GLU 18 10.505 42.471 94.883 1.00 89.02 0 ATOM 129 CG GLU 1810.769 43.929 94.547 1.00 97.06 0 ATOM 130 CD GLU 18 10.677 44.23993.069 1.00 39.46 0 ATOM 131 OE1 GLU 18 11.030 43.351 92.256 1.00 39.460 ATOM 132 OE2 GLU 18 10.265 45.375 92.727 1.00 39.46 0 ATOM 133 C GLU18 12.199 42.724 96.687 1.00 70.82 0 ATOM 134 O GLU 18 13.244 42.24996.240 1.00 88.34 0 ATOM 135 N VAL 19 12.172 43.793 97.480 1.00 28.97 0ATOM 137 CA VAL 19 13.394 44.470 97.891 1.00 28.97 0 ATOM 138 CB VAL 1913.139 45.968 96.207 1.00 6.66 0 ATOM 139 CG1 VAL 19 12.746 46.70296.942 1.00 6.86 0 ATOM 140 CG2 VAL 19 12.044 46.117 99.231 1.00 4.03 0ATOM 141 C VAL 19 14.079 43.805 99.081 1.00 28.97 0 ATOM 142 O VAL 1915.134 44.258 99.514 1.00 13.23 0 ATOM 143 N ARG 20 13.490 42.736 99.6151.00 2.00 0 ATOM 145 CA ARG 20 14.093 42.016 100.748 1.00 2.00 0 ATOM146 CB ARG 20 13.242 40.812 101.142 1.00 17.66 0 ATOM 147 CG ARG 2012.043 41.138 101.990 1.00 16.46 0 ATOM 148 CD ARG 20 11.192 39.899102.195 1.00 21.30 0 ATOM 149 NE ARG 20 12.006 38.733 102.532 1.00 18.450 ATOM 151 CZ ARG 20 11.559 37.481 102.546 1.00 23.19 0 ATOM 152 NH1 ARG20 10.288 37.232 102.249 1.00 28.11 0 ATOM 155 NH2 ARG 20 12.383 36.477102.836 1.00 21.96 0 ATOM 158 C ARG 20 15.480 41.521 100.333 1.00 2.00 0ATOM 159 O ARG 20 15.609 40.778 99.353 1.00 20.52 0 ATOM 160 N GLY 2116.514 41.956 101.047 1.00 61.97 0 ATOM 162 CA GLY 21 17.863 41.527100.718 1.00 65.49 0 ATOM 163 C GLY 21 18.702 42.522 99.930 1.00 66.05 0ATOM 164 O GLY 21 19.933 42.409 99.889 1.00 13.74 0 ATOM 165 N SER 2218.055 43.490 99.290 1.00 19.66 0 ATOM 167 CA SER 22 18.790 44.49198.523 1.00 17.08 0 ATOM 168 CB SER 22 17.874 45.159 97.481 1.00 26.61 0ATOM 169 OG SER 22 16.821 45.908 98.074 1.00 32.17 0 ATOM 171 C SER 2219.371 45.538 99.466 1.00 17.54 0 ATOM 172 O SER 22 19.047 45.558100.657 1.00 21.05 0 ATOM 173 N LYS 23 20.222 46.409 98.935 1.00 53.24 0ATOM 175 CA LYS 23 20.828 47.458 99.740 1.00 49.50 0 ATOM 176 CB LYS 2321.565 48.471 98.852 1.00 94.25 0 ATOM 177 CG LYS 23 20.639 49.42298.085 1.00 59.71 0 ATOM 178 CD LYS 23 21.341 50.716 97.688 1.00 94.25 0ATOM 179 CE LYS 23 20.346 51.775 97.214 1.00 59.77 0 ATOM 180 NZ LYS 2319.448 52.240 98.306 1.00 59.94 0 ATOM 184 C LYS 23 19.739 48.190100.528 1.00 49.47 0 ATOM 185 O LYS 23 18.659 48.488 99.998 1.00 59.94 0ATOM 186 N PRO 24 19.990 48.458 101.813 1.00 37.72 0 ATOM 187 CD PRO 2421.185 48.148 102.614 1.00 2.00 0 ATOM 188 CA PRO 24 18.987 49.165102.613 1.00 36.73 0 ATOM 189 CB PRO 24 19.618 49.217 104.004 1.00 2.000 ATOM 190 CG PRO 24 21.109 49.208 103.704 1.00 2.00 0 ATOM 191 C PRO 2418.798 50.551 102.019 1.00 33.26 0 ATOM 192 O PRO 24 19.752 51.325101.938 1.00 2.00 0 ATOM 193 N GLY 25 17.579 50.835 101.575 1.00 22.79 0ATOM 195 CA GLY 25 17.275 52.119 100.981 1.00 22.84 0 ATOM 196 C GLY 2516.653 51.904 99.624 1.00 18.00 0 ATOM 197 O GLY 25 16.098 52.827 99.0371.00 28.24 0 ATOM 198 N LYS 26 16.750 50.679 99.116 1.00 2.00 0 ATOM 200CA LYS 26 16.174 50.351 97.817 1.00 2.00 0 ATOM 201 CB LYS 26 16.46948.892 97.458 1.00 45.45 0 ATOM 202 CG LYS 26 15.931 48.437 96.110 1.0055.13 0 ATOM 203 CD LYS 26 16.209 49.435 94.979 1.00 59.31 0 ATOM 204 CELYS 26 17.694 49.644 94.691 1.00 60.93 0 ATOM 205 NZ LYS 26 17.88350.619 93.569 1.00 68.30 0 ATOM 209 C LYS 26 14.674 50.624 97.856 1.002.00 0 ATOM 210 O LYS 26 13.916 49.964 98.566 1.00 42.74 0 ATOM 211 NASN 27 14.278 51.648 97.111 1.00 2.00 0 ATOM 213 CA ASN 27 12.894 52.08697.027 1.00 2.00 0 ATOM 214 CB ASN 27 12.836 53.526 96.517 1.00 50.37 0ATOM 215 CG ASN 27 13.257 54.525 97.563 1.00 56.29 0 ATOM 216 OD1 ASN 2712.929 54.381 98.740 1.00 61.45 0 ATOM 217 ND2 ASN 27 13.982 55.55197.142 1.00 59.50 0 ATOM 220 C ASN 27 11.964 51.219 96.183 1.00 2.00 0ATOM 221 O ASN 27 12.384 50.256 95.540 1.00 54.11 0 ATOM 222 N VAL 2810.689 51.590 96.209 1.00 12.34 0 ATOM 224 CA VAL 28 9.646 50.910 95.4731.00 12.71 0 ATOM 225 CB1 VAL 28 9.126 49.693 96.283 1.00 2.00 0 ATOM226 CG1 VAL 28 8.777 50.111 97.684 1.00 2.00 0 ATOM 227 CG2 VAL 28 7.93249.053 95.599 1.00 2.00 0 ATOM 228 C VAL 28 8.549 51.935 95.145 1.0019.22 0 ATOM 229 O VAL 28 7.757 52.363 96.000 1.00 2.00 0 ATOM 230 N GLN29 8.548 52.372 93.892 1.00 26.36 0 ATOM 232 CA GLN 29 7.586 53.36593.424 1.00 27.97 0 ATOM 233 CB GLN 29 8.203 54.239 92.325 1.00 11.00 0ATOM 234 CG GLN 29 7.479 55.543 92.080 1.00 8.47 0 ATOM 235 CD GLN 297.684 56.541 93.201 1.00 11.72 0 ATOM 236 OE1 GLN 29 7.097 57.619 93.1981.00 13.39 0 ATOM 237 NE2 GLN 29 8.525 56.195 94.159 1.00 12.03 0 ATOM240 C GLN 29 6.347 52.688 92.887 1.00 27.73 0 ATOM 241 O GLN 29 6.40151.934 91.926 1.00 8.22 0 ATOM 242 N LEU 30 5.229 52.941 93.531 1.002.00 0 ATOM 244 CA LEU 30 3.978 52.359 93.087 1.00 2.00 0 ATOM 245 CBLEU 30 3.157 51.859 94.279 1.00 18.06 0 ATOM 246 CG LEU 30 3.381 50.41694.729 1.00 18.06 0 ATOM 247 CD1 LEU 30 4.857 50.152 94.928 1.00 18.06 0ATOM 248 CD2 LEU 30 2.603 50.171 96.009 1.00 18.06 0 ATOM 249 C LEU 303.223 53.441 92.348 1.00 2.00 0 ATOM 250 O LEU 30 3.363 54.621 92.6641.00 18.06 0 ATOM 251 N GLN 31 2.441 53.050 91.355 1.00 75.78 0 ATOM 253CA GLN 31 1.679 54.026 90.599 1.00 80.15 0 ATOM 254 CB GLN 31 0.78253.336 89.593 1.00 2.00 0 ATOM 255 CG GLN 31 1.448 52.204 68.883 1.002.00 0 ATOM 256 CD GLN 31 0.498 51.469 87.976 1.00 2.00 0 ATOM 257 OE1GLN 31 0.933 50.694 87.122 1.00 2.00 0 ATOM 258 NE2 GLN 31 −0.809 51.69888.150 1.00 2.00 0 ATOM 261 C GLN 31 0.819 54.783 91.585 1.00 79.69 0ATOM 262 O GLN 31 0.276 54.187 92.512 1.00 2.00 0 ATOM 263 N GLU 320.716 56.091 91.380 1.00 2.00 0 ATOM 265 CA GLU 32 −0.074 56.979 92.2281.00 2.00 0 ATOM 266 CB GLU 32 −0.236 58.333 91.523 1.00 57.69 0 ATOM267 CG GLU 32 −1.206 59.320 92.181 1.00 62.36 0 ATOM 268 CD GLU 32−1.652 60.426 91.226 1.00 60.46 0 ATOM 269 OE1 GLU 32 −2.596 61.17691.570 1.00 59.61 0 ATOM 270 OE2 GLU 32 −1.059 60.543 90.128 1.00 62.850 ATOM 271 C GLU 32 −1.449 56.370 92.539 1.00 2.00 0 ATOM 272 O GLU 32−1.875 56.341 93.695 1.00 53.86 0 ATOM 273 N ASN 33 −2.127 55.865 91.5171.00 6.25 0 ATOM 275 CA ASN 33 −3.445 55.278 91.714 1.00 7.45 0 ATOM 276CB ASN 33 −4.134 55.026 90.364 1.00 30.19 0 ATOM 277 CG ASN 33 −3.29154.199 89.412 1.00 30.49 0 ATOM 278 OD1 ASN 33 −2.592 53.261 89.810 1.0032.73 0 ATOM 279 ND2 ASN 33 −3.349 54.551 88.141 1.00 33.00 0 ATOM 282 CASN 33 −3.448 54.002 92.565 1.00 10.76 0 ATOM 283 O ASN 33 −4.466 53.67093.182 1.00 23.15 0 ATOM 284 N GLU 34 −2.322 53.292 92.598 1.00 27.56 0ATOM 286 CA GLU 34 −2.217 52.081 93.402 1.00 24.54 0 ATOM 287 CB GLU 34−1.005 51.251 92.985 1.00 23.44 0 ATOM 288 CG GLU 34 −1.203 50.47991.698 1.00 22.75 0 ATOM 289 CD GLU 34 −0.120 49.438 91.479 1.00 28.72 0ATOM 290 OE1 GLU 34 −0.443 48.226 91.460 1.00 34.33 0 ATOM 291 OE2 GLU34 1.055 49.834 91.330 1.00 31.11 0 ATOM 292 C GLU 34 −2.106 52.47094.871 1.00 25.10 0 ATOM 293 O GLU 34 −2.797 51.923 95.716 1.00 19.23 0ATOM 294 N ILE 35 −1.244 53.430 95.172 1.00 14.94 0 ATOM 296 CA ILE 35−1.083 53.889 96.541 1.00 28.22 0 ATOM 297 CB ILE 35 0.045 54.888 96.6321.00 2.00 0 ATOM 298 CG2 ILE 35 0.013 55.607 97.961 1.00 2.00 0 ATOM 299CG1 ILE 35 1.354 54.154 96.395 1.00 2.00 0 ATOM 300 CD1 ILE 35 2.55455.045 96.414 1.00 2.00 0 ATOM 301 C ILE 35 −2.368 54.536 97.034 1.0013.21 0 ATOM 302 O ILE 35 −2.794 54.302 98.172 1.00 2.00 0 ATOM 303 NARG 36 −2.985 55.340 96.171 1.00 2.00 0 ATOM 305 CA ARG 36 −4.237 56.01596.506 1.00 2.00 0 ATOM 306 CB ARG 36 −4.578 57.082 95.446 1.00 8.06 0ATOM 307 CG ARG 36 −5.725 56.703 94.502 1.00 14.08 0 ATOM 308 CD ARG 36−6.059 57.824 93.555 1.00 20.55 0 ATOM 309 NE ARG 36 −6.465 59.02994.267 1.00 14.43 0 ATOM 311 CZ ARG 36 −7.726 59.383 94.491 1.00 22.93 0ATOM 312 NH1 ARG 36 −8.729 58.618 94.064 1.00 22.80 0 ATOM 315 NH2 ARG36 −7.987 60.518 95.134 1.00 19.87 0 ATOM 318 C ARG 36 −5.394 55.00596.657 1.00 2.00 0 ATOM 319 O ARG 36 −6.415 55.305 97.279 1.00 2.00 0ATOM 320 N GLY 37 −5.240 53.828 96.059 1.00 2.00 0 ATOM 322 CA GLY 37−6.264 52.814 96.177 1.00 2.00 0 ATOM 323 C GLY 37 −6.118 52.251 97.5751.00 2.00 0 ATOM 324 O GLY 37 −7.106 52.030 98.289 1.00 2.00 0 ATOM 325N LEU 38 −4.864 52.039 97.975 1.00 8.88 0 ATOM 327 CA LEU 38 −4.55851.515 99.288 1.00 8.88 0 ATOM 328 CB LEU 38 −3.061 51.550 99.559 1.002.00 0 ATOM 329 CG LEU 38 −2.202 50.559 98.784 1.00 2.00 0 ATOM 330 CD1LEU 38 −0.765 50.804 99.171 1.00 2.00 0 ATOM 331 CD2 LEU 38 −2.61449.106 99.063 1.00 2.00 0 ATOM 332 C LEU 38 −5.278 52.406 100.258 1.008.88 0 ATOM 333 O LEU 38 −6.130 51.928 101.002 1.00 2.00 0 ATOM 334 NCYS 39 −4.976 53.705 100.216 1.00 2.00 0 ATOM 336 CA CYS 39 −5.61354.690 101.099 1.00 2.00 0 ATOM 337 CB CYS 39 −5.339 56.096 100.607 1.002.00 0 ATOM 338 SG CYS 39 −3.719 56.686 100.830 1.00 2.00 0 ATOM 339 CCYS 39 −7.140 54.555 101.201 1.00 2.00 0 ATOM 340 O CYS 39 −7.708 54.580102.296 1.00 2.00 0 ATOM 341 N LEU 40 −7.793 54.412 100.053 1.00 2.00 0ATOM 343 CA LEU 40 −9.233 54.330 99.999 1.00 2.00 0 ATOM 344 CB LEU 40−9.735 54.704 98.599 1.00 2.00 0 ATOM 345 CG LEU 40 −10.127 56.18498.419 1.00 2.00 0 ATOM 346 CD1 LEU 40 −9.020 57.095 98.939 1.00 2.00 0ATOM 347 CD2 LEU 40 −10.401 56.484 96.960 1.00 2.00 0 ATOM 348 C LEU 40−9.817 53.014 100.426 1.00 2.00 0 ATOM 349 O LEU 40 −10.853 52.987101.071 1.00 2.00 0 ATOM 350 N LYS 41 −9.174 51.916 100.081 1.00 7.31 0ATOM 352 CA LYS 41 −9.721 50.630 100.468 1.00 7.31 0 ATOM 353 CB LYS 41−9.123 49.517 99.598 1.00 2.00 0 ATOM 354 CG LYS 41 −9.685 48.157 99.8841.00 2.00 0 ATOM 355 CD LYS 41 −11.188 48.216 99.923 1.00 2.00 0 ATOM356 CE LYS 41 −11.746 46.968 100.560 1.00 2.00 0 ATOM 357 NZ LYS 41−11.188 45.721 99.910 1.00 2.00 0 ATOM 361 C LYS 41 9.475 50.376 101.9631.00 7.31 0 ATOM 362 O LYS 41 −10.375 49.934 102.674 1.00 2.00 0 ATOM363 N SER 42 −8.266 50.666 102.440 1.00 3.34 0 ATOM 365 CA SER 42 −7.93550.477 103.844 1.00 3.34 0 ATOM 366 CB SER 42 −6.496 50.854 104.094 1.002.00 0 ATOM 367 OG SER 42 −6.315 52.215 103.766 1.00 2.00 0 ATOM 369 CSER 42 −8.844 51.398 104.650 1.00 7.34 0 ATOM 370 O SER 42 −9.504 50.955105.600 1.00 2.00 0 ATOM 371 N ARG 43 −8.906 52.671 104.247 1.00 2.00 0ATOM 373 CA ARG 43 −9.746 53.650 104.928 1.00 2.00 0 ATOM 374 CB ARG 43−9.856 54.946 104.135 1.00 2.00 0 ATOM 375 CG ARG 43 −10.530 56.063104.917 1.00 2.00 0 ATOM 376 CD ARG 43 −11.541 56.826 104.097 1.00 2.000 ATOM 377 NE ARG 43 −11.803 58.137 104.679 1.00 2.00 0 ATOM 379 CZ ARG43 −13.010 58.603 105.005 1.00 2.00 0 ATOM 380 NH1 ARG 43 −14.112 57.877104.826 1.00 2.00 0 ATOM 383 NH2 ARG 43 −13.120 59.827 105.502 1.00 2.000 ATOM 386 C ARG 43 −11.136 53.103 105.104 1.00 2.00 0 ATOM 387 O ARG 43−11.800 53.385 106.083 1.00 2.00 0 ATOM 388 N GLU 44 −11.574 52.321104.130 1.00 19.11 0 ATOM 390 CA GLU 44 −12.901 51.727 104.146 1.0019.41 0 ATOM 391 CB GLU 44 −13.179 51.107 102.776 1.00 23.41 0 ATOM 392CG GLU 44 −14.599 50.695 102.542 1.00 34.12 0 ATOM 393 CD GLU 44 −14.71149.574 101.532 1.00 38.72 0 ATOM 394 OE1 GLU 44 −15.634 49.626 100.6991.00 43.51 0 ATOM 395 OE2 GLU 44 −13.886 48.639 101.576 1.00 35.90 0ATOM 396 C GLU 44 −12.987 50.674 105.260 1.00 20.56 0 ATOM 397 O GLU 44−13.967 50.622 106.007 1.00 24.61 0 ATOM 398 N ILE 45 −11.945 49.857105.379 1.00 8.30 0 ATOM 400 CA ILE 45 −11.896 48.811 106.384 1.00 8.300 ATOM 401 CB ILE 45 −10.720 47.876 106.133 1.00 2.00 0 ATOM 402 CG2 ILE45 −10.795 46.678 107.049 1.00 2.00 0 ATOM 403 CG1 ILE 45 −10.774 47.382104.698 1.00 2.00 0 ATOM 404 CD1 ILE 45 −9.559 46.593 104.285 1.00 2.000 ATOM 405 C ILE 45 −11.775 49.401 107.784 1.00 8.30 0 ATOM 406 O ILE 45−12.249 48.819 108.756 1.00 2.00 0 ATOM 407 N PHE 46 −11.134 50.550107.905 1.00 2.00 0 ATOM 409 CA PHE 46 −11.013 51.154 109.214 1.00 2.000 ATOM 410 CB PHE 46 −10.076 52.352 109.176 1.00 2.00 0 ATOM 411 CG PHE46 −8.665 51.992 108.843 1.00 2.00 0 ATOM 412 CD1 PHE 46 −7.829 52.906108.223 1.00 2.00 0 ATOM 413 CD2 PHE 46 −8.168 50.726 109.151 1.00 2.000 ATOM 414 CE1 PHE 46 −6.516 52.558 107.915 1.00 2.00 0 ATOM 415 CE2 PHE46 −6.859 50.375 108.845 1.00 2.00 0 ATOM 416 CZ PHE 46 −6.027 51.289108.227 1.00 2.00 0 ATOM 417 C PHE 46 −12.396 51.561 109.686 1.00 2.00 0ATOM 418 O PHE 46 −12.840 51.109 110.731 1.00 2.00 0 ATOM 419 N LEU 47−13.106 52.358 108.896 1.00 2.00 0 ATOM 421 CA LEU 47 −14.441 52.807109.277 1.00 2.00 0 ATOM 422 CB LEU 47 −15.043 53.705 108.190 1.00 2.000 ATOM 423 CG LEU 47 −14.641 55.181 108.110 1.00 2.00 0 ATOM 424 CD1 LEU47 −14.094 55.641 109.462 1.00 2.00 0 ATOM 425 CD2 LEU 47 −13.619 55.397107.030 1.00 2.00 0 ATOM 426 C LEU 47 −15.424 51.677 109.578 1.00 2.00 0ATOM 427 O LEU 47 −16.432 51.905 110.243 1.00 2.00 0 ATOM 428 N SER 48−15.145 50.469 109.091 1.00 2.00 0 ATOM 430 CA SER 48 −16.037 49.339109.305 1.00 2.00 0 ATOM 431 CB SER 48 −16.121 48.483 108.039 1.00 23.350 ATOM 432 OG SER 48 −14.844 48.045 107.622 1.00 30.63 0 ATOM 434 C SER48 −15.624 48.473 110.482 1.00 2.00 0 ATOM 435 O SER 48 −16.184 47.390110.700 1.00 28.70 0 ATOM 436 N GLN 49 −14.627 48.922 111.225 1.00 61.720 ATOM 438 CA GLN 49 −14.175 48.193 112.394 1.00 63.25 0 ATOM 439 CB GLN49 −12.763 47.645 112.173 1.00 13.32 0 ATOM 440 CG GLN 49 −12.668 46.511111.146 1.00 7.00 0 ATOM 441 CD GLN 49 −11.246 45.952 110.995 1.00 8.880 ATOM 442 OE1 GLN 49 −10.251 46.640 111.258 1.00 9.23 0 ATOM 443 NE2GLN 49 −11.151 44.700 110.573 1.00 7.45 0 ATOM 446 C GLN 49 −14.21049.205 113.540 1.00 63.04 0 ATOM 447 O GLN 49 −13.864 50.373 113.3571.00 14.61 0 ATOM 448 N PRO 50 −14.645 48.778 114.734 1.00 34.58 0 ATOM449 CD PRO 50 −14.967 47.399 115.118 1.00 4.65 0 ATOM 450 CA PRO 50−14.728 49.657 115.901 1.00 34.58 0 ATOM 451 CB PRO 50 −15.037 48.678117.032 1.00 2.00 0 ATOM 452 CG PRO 50 −14.496 47.365 116.531 1.00 2.000 ATOM 453 C PRO 50 −13.459 50.468 116.164 1.00 34.58 0 ATOM 454 O PRO50 −12.356 50.002 115.884 1.00 2.95 0 ATOM 455 N ILE 51 −13.626 51.673116.708 1.00 2.00 0 ATOM 457 CA ILE 51 −12.499 52.550 117.015 1.00 2.000 ATOM 458 CB ILE 51 −13.001 53.975 117.318 1.00 2.00 0 ATOM 459 CG2 ILE51 −13.642 54.035 118.673 1.00 2.00 0 ATOM 460 CG1 ILE 51 −11.850 54.958117.295 1.00 2.00 0 ATOM 461 CD1 ILE 51 −12.328 56.370 117.351 1.00 2.000 ATOM 462 C ILE 51 −11.650 51.996 118.172 1.00 2.00 0 ATOM 463 O ILE 51−10.456 52.303 118.283 1.00 2.00 0 ATOM 464 N LEU 52 −12.284 51.187119.028 1.00 2.00 0 ATOM 466 CA LEU 52 −11.622 50.522 120.159 1.00 2.000 ATOM 467 CB LEU 52 −12.391 50.746 121.473 1.00 2.00 0 ATOM 468 CG LEU52 −11.655 50.518 122.808 1.00 2.00 0 ATOM 469 CD1 LEU 52 −12.559 50.899123.944 1.00 2.00 0 ATOM 470 CD2 LEU 52 −11.233 49.085 122.973 1.00 2.000 ATOM 471 C LEU 52 −11.660 49.033 119.770 1.00 2.00 0 ATOM 472 O LEU 52−12.652 48.331 120.006 1.00 2.00 0 ATOM 473 N LEU 53 −10.584 48.576119.136 1.00 2.00 0 ATOM 475 CA LEU 53 −10.464 47.204 118.667 1.00 2.000 ATOM 476 CB LEU 53 −9.066 46.982 118.069 1.00 2.00 0 ATOM 477 CG LEU53 −8.802 47.038 116.555 1.00 2.00 0 ATOM 478 CD1 LEU 53 −9.835 47.889115.810 1.00 2.00 0 ATOM 479 CD2 LEU 53 −7.401 47.550 116.353 1.00 2.000 ATOM 480 C LEU 53 −10.686 46.230 119.792 1.00 2.00 0 ATOM 481 O LEU 53−10.365 46.522 120.937 1.00 2.00 0 ATOM 482 N GLU 54 −11.251 45.076119.472 1.00 19.77 0 ATOM 484 CA GLU 54 −11.465 44.049 120.474 1.0020.13 0 ATOM 485 CB GLU 54 −12.955 43.809 120.714 1.00 56.32 0 ATOM 486CG GLU 54 −13.244 43.362 122.135 1.00 66.19 0 ATOM 487 CD GLU 54 −14.66842.873 122.346 1.00 69.64 0 ATOM 488 OE1 GLU 54 −15.613 43.627 122.0241.00 78.06 0 ATOM 489 OE2 GLU 54 −14.839 41.734 122.848 1.00 73.27 0ATOM 490 C GLU 54 −10.798 42.820 119.882 1.00 19.77 0 ATOM 491 O GLU 54−11.451 41.949 119.313 1.00 48.57 0 ATOM 492 N LEU 55 −9.473 42.800119.974 1.00 2.00 0 ATOM 494 CA LEU 55 −8.666 41.712 119.452 1.00 2.00 0ATOM 495 CB LEU 55 −7.245 42.188 119.176 1.00 2.00 0 ATOM 496 CG LEU 55−7.148 43.458 118.336 1.00 2.00 0 ATOM 497 CD1 LEU 55 −5.695 43.728117.961 1.00 2.00 0 ATOM 498 CD2 LEU 55 −8.024 43.298 117.108 1.00 2.000 ATOM 499 C LEU 55 −8.624 40.586 120.456 1.00 2.00 0 ATOM 500 O LEU 55−8.724 40.807 121.664 1.00 10.45 0 ATOM 501 N GLU 56 −8.448 39.374119.961 1.00 2.00 0 ATOM 503 CA GLU 56 −8.407 38.224 120.826 1.00 2.00 0ATOM 504 CB GLU 56 −9.741 37.467 120.729 1.00 18.79 0 ATOM 505 CG GLU 56−10.989 38.283 121.171 1.00 30.34 0 ATOM 506 CD GLU 56 −11.012 38.677122.658 1.00 42.94 0 ATOM 507 OE1 GLU 56 −11.188 37.789 123.526 1.0044.70 0 ATOM 508 OE2 GLU 56 −10.872 39.882 122.955 1.00 47.48 0 ATOM 509C GLU 56 −7.244 37.343 120.393 1.00 2.00 0 ATOM 510 O GLU 56 −6.29237.839 119.800 1.00 3.40 0 ATOM 511 N ALA 57 −7.312 36.062 120.765 1.0065.90 0 ATOM 513 CA ALA 57 −6.336 35.026 120.405 1.00 62.88 0 ATOM 514CB ALA 57 −6.750 34.434 119.039 1.00 26.61 0 ATOM 515 C ALA 57 −4.86735.474 120.387 1.00 68.03 0 ATOM 516 O ALA 57 −4.543 36.519 120.932 1.0014.05 0 ATOM 517 N PRO 58 −3.951 34.632 119.857 1.00 2.00 0 ATOM 518 CDPRO 58 −4.109 33.203 119.533 1.00 9.23 0 ATOM 519 CA PRO 58 −2.52634.988 119.782 1.00 2.00 0 ATOM 520 CB PRO 58 −1.852 33.645 119.501 1.009.23 0 ATOM 521 CG PRO 58 −2.881 32.926 118.711 1.00 9.49 0 ATOM 522 CPRO 58 −2.176 36.002 118.672 1.00 2.00 0 ATOM 523 O PRO 58 −2.688 35.901117.540 1.00 12.29 0 ATOM 524 N LEU 59 −1.284 36.947 118.975 1.00 17.050 ATOM 526 CA LEU 59 −0.856 37.944 117.989 1.00 17.05 0 ATOM 527 CB LEU59 −1.862 39.100 117.895 1.00 2.00 0 ATOM 528 CG LEU 59 −1.842 40.166118.993 1.00 2.00 0 ATOM 529 CD1 LEU 59 −2.880 41.252 118.706 1.00 2.000 ATOM 530 CD2 LEU 59 −2.117 39.498 120.339 1.00 2.00 0 ATOM 531 C LEU59 0.510 38.501 118.351 1.00 17.05 0 ATOM 532 O LEU 59 1.030 38.229119.429 1.00 2.00 0 ATOM 533 N LYS 60 1.103 39.259 117.438 1.00 21.64 0ATOM 535 CA LYS 60 2.399 39.872 117.688 1.00 21.64 0 ATOM 536 CB LYS 603.358 39.655 116.513 1.00 10.44 0 ATOM 537 CG LYS 60 3.185 38.327115.768 1.00 11.70 0 ATOM 538 CD LYS 60 3.556 37.101 116.608 1.00 12.190 ATOM 539 CE LYS 60 5.034 36.783 116.544 1.00 8.37 0 ATOM 540 NZ LYS 605.824 37.927 117.066 1.00 8.37 0 ATOM 544 C LYS 60 2.063 41.353 117.8041.00 21.64 0 ATOM 545 O LYS 60 1.142 41.832 117.128 1.00 13.47 0 ATOM546 N ILE 61 2.757 42.072 118.680 1.00 20.60 0 ATOM 548 CA ILE 61 2.49943.496 118.822 1.00 22.32 0 ATOM 549 CB ILE 61 2.032 43.859 120.240 1.002.00 0 ATOM 550 CG2 ILE 61 1.485 45.288 120.239 1.00 2.00 0 ATOM 551 CG1ILE 61 0.940 42.881 120.702 1.00 2.00 0 ATOM 552 CD1 ILE 61 0.019 43.422121.783 1.00 2.00 0 ATOM 553 C ILE 61 3.791 44.220 118.494 1.00 23.52 0ATOM 554 O ILE 61 4.862 43.752 118.868 1.00 2.00 0 ATOM 555 N CYS 623.698 45.341 117.787 1.00 2.00 0 ATOM 557 CA CYS 62 4.874 46.095 117.3931.00 2.00 0 ATOM 558 CB CYS 62 5.147 45.925 115.895 1.00 11.47 0 ATOM559 SG CYS 62 5.439 44.252 115.266 1.00 11.47 0 ATOM 560 C CYS 62 4.60747.560 117.658 1.00 2.00 0 ATOM 561 O CYS 62 3.451 47.957 117.751 1.0011.47 0 ATOM 562 N GLY 63 5.662 48.367 117.739 1.00 2.00 0 ATOM 564 CAGLY 63 5.507 49.789 117.971 1.00 2.00 0 ATOM 565 C GLY 63 6.548 50.610117.222 1.00 2.00 0 ATOM 566 O GLY 63 7.624 50.109 116.897 1.00 11.39 0ATOM 567 N ASP 64 6.216 51.870 116.959 1.00 2.00 0 ATOM 569 CA ASP 647.068 52.834 116.255 1.00 3.88 0 ATOM 570 CB ASP 64 7.805 53.721 117.2451.00 6.62 0 ATOM 571 CG ASP 64 6.873 54.589 118.025 1.00 6.62 0 ATOM 572OD1 ASP 64 5.996 54.019 118.708 1.00 12.70 0 ATOM 573 OD2 ASP 64 7.00355.829 117.946 1.00 12.70 0 ATOM 574 C ASP 64 8.064 52.362 115.221 1.0014.37 0 ATOM 575 O ASP 64 9.221 52.080 115.557 1.00 8.57 0 ATOM 576 NILE 65 7.619 52.326 113.963 1.00 5.19 0 ATOM 578 CA ILE 65 8.471 51.924112.849 1.00 3.07 0 ATOM 579 CB ILE 65 7.663 51.310 111.672 1.00 2.00 0ATOM 580 CG2 ILE 65 8.609 50.903 110.535 1.00 2.00 0 ATOM 581 CG1 ILE 656.871 50.095 112.151 1.00 2.00 0 ATOM 582 CD1 ILE 65 7.723 49.023112.763 1.00 2.00 0 ATOM 583 C ILE 65 9.185 53.177 112.357 1.00 2.24 0ATOM 584 O ILE 65 10.379 53.148 112.101 1.00 2.00 0 ATOM 585 N HIS 668.451 54.276 112.235 1.00 2.00 0 ATOM 587 CA HIS 66 9.022 55.536 111.7761.00 2.00 0 ATOM 588 C HIS 66 9.847 55.525 110.502 1.00 2.00 0 ATOM 589O HIS 66 11.042 55.818 110.539 1.00 12.38 0 ATOM 590 CB HIS 66 9.84656.182 112.88 1.00 8.25 0 ATOM 591 CG HIS 66 9.040 57.046 113.782 1.008.25 0 ATOM 592 ND1 HIS 66 8.104 57.949 113.337 1.00 8.25 0 ATOM 594 CD2HIS 66 9.001 57.109 115.133 1.00 8.25 0 ATOM 595 NE2 HIS 66 8.042 58.043115.526 1.00 6.25 0 ATOM 596 CE1 HIS 66 7.536 58.516 114.399 1.00 8.25 0ATOM 597 N GLY 67 9.197 55.195 109.385 1.00 11.52 0 ATOM 599 CA GLY 679.835 55.174 108.078 1.00 11.52 0 ATOM 600 C GLY 67 11.029 54.266107.868 1.00 11.52 0 ATOM 601 O GLY 67 11.782 54.462 106.915 1.00 81.940 ATOM 602 N GLN 68 11.214 53.272 108.730 1.00 18.96 0 ATOM 604 CA GLN68 12.353 52.364 108.602 1.00 20.32 0 ATOM 605 CB GLN 68 13.007 52.139109.957 1.00 6.39 0 ATOM 606 CG GLN 68 13.261 53.407 110.711 1.00 4.06 0ATOM 607 CD GLN 68 14.315 53.239 111.757 1.00 7.01 0 ATOM 608 OE1 GLN 6815.013 54.197 112.111 1.00 8.21 0 ATOM 609 NE2 GLN 68 14.453 52.020112.270 1.00 5.84 0 ATOM 612 C GLN 68 11.916 51.039 108.010 1.00 18.72 0ATOM 613 O GLN 68 12.018 49.975 108.634 1.00 10.73 0 ATOM 614 N TYR 6911.450 51.119 106.777 1.00 5.70 0 ATOM 616 CA TYR 69 10.959 49.970106.054 1.00 5.52 0 ATOM 617 CB TYR 69 10.807 50.313 104.587 1.00 2.00 0ATOM 618 CG TYR 69 9.988 49.322 103.841 1.00 2.00 0 ATOM 619 CD1 TYR 698.658 49.099 104.187 1.00 2.00 0 ATOM 620 CE1 TYR 69 7.873 48.194103.477 1.00 2.00 0 ATOM 621 CD2 TYR 69 10.526 48.611 102.762 1.00 2.000 ATOM 622 CE2 TYR 69 9.751 47.701 102.038 1.00 2.00 0 ATOM 623 CZ TYR69 8.431 47.506 102.408 1.00 2.00 0 ATOM 624 OH TYR 69 7.656 46.632101.715 1.00 2.00 0 ATOM 626 C TYR 69 11.815 48.730 106.188 1.00 8.52 0ATOM 627 O TYR 69 11.284 47.641 106.393 1.00 2.00 0 ATOM 628 N TYR 7013.132 48.877 106.093 1.00 2.00 0 ATOM 630 CA TYR 70 13.992 47.703106.186 1.00 2.00 0 ATOM 631 CB TYR 70 15.420 48.031 105.741 1.00 64.800 ATOM 632 CG TYR 70 15.533 47.964 104.231 1.00 70.91 0 ATOM 633 CD1 TYR70 15.026 48.993 103.431 1.00 73.06 0 ATOM 634 CE1 TYR 70 15.082 48.924102.041 1.00 68.38 0 ATOM 635 CD2 TYR 70 16.107 46.857 103.597 1.0073.86 0 ATOM 636 CE2 TYR 70 16.171 46.780 102.200 1.00 70.94 0 ATOM 637CZ TYR 70 15.654 47.820 101.433 1.00 72.52 0 ATOM 638 OH TYR 70 15.71247.779 100.062 1.00 70.35 0 ATOM 640 C TYR 70 13.943 47.007 107.538 1.002.00 0 ATOM 641 O TYR 70 13.967 45.771 107.610 1.00 64.08 0 ATOM 642 NASP 71 13.821 47.789 108.608 1.00 7.87 0 ATOM 644 CA ASP 71 13.72847.203 109.927 1.00 6.73 0 ATOM 645 CB ASP 71 14.030 48.242 110.998 1.0014.88 0 ATOM 646 CG ASP 71 15.514 48.600 111.054 1.00 25.87 0 ATOM 647OD1 ASP 71 15.826 49.778 111.293 1.00 24.58 0 ATOM 648 OD2 ASP 71 16.37547.714 110.861 1.00 21.52 0 ATOM 649 C ASP 71 12.331 46.622 110.051 1.006.73 0 ATOM 650 O ASP 71 12.116 45.697 110.826 1.00 14.14 0 ATOM 651 NLEU 72 11.399 47.148 109.250 1.00 2.00 0 ATOM 653 CA LEU 72 10.01546.654 109.208 1.00 2.00 0 ATOM 654 CB LEU 72 9.094 47.613 108.456 1.002.00 0 ATOM 655 CG LEU 72 7.771 46.946 108.067 1.00 2.00 0 ATOM 656 CD1LEU 72 7.025 46.562 109.342 1.00 2.00 0 ATOM 657 CD2 LEU 72 6.935 47.867107.208 1.00 2.00 0 ATOM 658 C LEU 72 10.014 45.320 108.476 1.00 2.00 0ATOM 659 O LEU 72 9.259 44.401 108.814 1.00 2.00 0 ATOM 660 N LEU 7310.848 45.231 107.449 1.00 44.35 0 ATOM 662 CA LEU 73 10.968 44.005106.693 1.00 42.59 0 ATOM 663 CB LEU 73 11.846 44.218 105.460 1.00 2.000 ATOM 664 CG LEU 73 11.248 44.759 104.160 1.00 2.00 0 ATOM 665 CD1 LEU73 12.324 44.728 103.101 1.00 2.00 0 ATOM 666 CD2 LEU 73 10.052 43.920103.725 1.00 2.00 0 ATOM 667 C LEU 73 11.603 42.978 107.629 1.00 42.42 0ATOM 668 O LEU 73 11.059 41.890 107.817 1.00 2.00 0 ATOM 669 N ARG 7412.729 43.354 108.241 1.00 2.00 0 ATOM 671 CA ARG 74 13.462 42.494109.179 1.00 2.00 0 ATOM 672 CB ARG 74 14.591 43.285 109.840 1.00 31.220 ATOM 673 CG ARG 74 15.909 43.478 108.958 1.00 31.18 0 ATOM 674 CD ARG74 16.944 44.146 109.710 1.00 37.63 0 ATOM 675 NE ARG 74 17.250 43.459110.960 1.00 37.44 0 ATOM 677 CZ ARG 74 17.210 44.041 112.157 1.00 41.100 ATOM 678 NH1 ARG 74 16.888 45.327 112.265 1.00 39.99 0 ATOM 681 NH2ARG 74 17.476 43.339 113.254 1.00 39.22 0 ATOM 684 C ARG 74 12.56941.896 110.259 1.00 2.00 0 ATOM 685 O ARG 74 12.621 40.703 110.544 1.0029.18 0 ATOM 686 N LEU 75 11.747 42.742 110.853 1.00 2.00 0 ATOM 688 CALEU 75 10.818 42.331 111.901 1.00 2.00 0 ATOM 689 CB LEU 75 10.06943.580 112.407 1.00 3.53 0 ATOM 690 CG LEU 75 8.704 43.654 113.091 1.003.53 0 ATOM 691 CD1 LEU 75 8.554 45.074 113.606 1.00 3.53 0 ATOM 692 CD2LEU 75 7.561 43.337 112.138 1.00 3.53 0 ATOM 693 C LEU 75 9.850 41.279111.384 1.00 2.00 0 ATOM 694 O LEU 75 9.594 40.284 112.043 1.00 3.53 0ATOM 695 N PHE 76 9.325 41.514 110.190 1.00 2.00 0 ATOM 697 CA PHE 768.382 40.604 109.573 1.00 2.00 0 ATOM 698 CB PHE 76 7.883 41.173 108.2551.00 2.00 0 ATOM 699 CG PHE 76 6.626 41.966 108.382 1.00 2.00 0 ATOM 700CD1 PHE 76 6.447 43.124 107.642 1.00 2.00 0 ATOM 701 CD2 PHE 76 5.61641.550 109.240 1.00 2.00 0 ATOM 702 CE1 PHE 76 5.277 43.852 107.753 1.002.00 0 ATOM 703 CE2 PHE 76 4.447 42.270 109.358 1.00 2.00 0 ATOM 704 CZPHE 76 4.272 43.423 108.615 1.00 2.00 0 ATOM 705 C PHE 76 9.016 39.271109.322 1.00 2.00 0 ATOM 706 O PHE 76 8.339 38.260 109.363 1.00 2.00 0ATOM 707 N GLU 77 10.316 39.283 109.043 1.00 2.00 0 ATOM 709 CA GLU 7711.080 38.065 108.789 1.00 2.00 0 ATOM 710 CB GLU 77 12.436 38.409108.191 1.00 84.85 0 ATOM 711 CG GLU 77 12.331 39.117 106.861 1.00 89.260 ATOM 712 CD GLU 77 13.651 39.690 106.391 1.00 98.60 0 ATOM 713 OE1 GLU77 13.627 40.493 105.434 1.00 98.04 0 ATOM 714 OE2 GLU 77 14.709 39.345106.973 1.00 0.97 0 ATOM 715 C GLU 77 11.262 37.316 110.100 1.00 2.00 0ATOM 716 O GLU 77 11.469 36.103 110.099 1.00 80.75 0 ATOM 717 N TYR 7811.182 38.045 111.216 1.00 15.29 0 ATOM 719 CA TYR 78 11.325 37.449112.538 1.00 15.29 0 ATOM 720 CB TYR 78 11.945 38.455 113.514 1.00 67.130 ATOM 721 CG TYR 78 12.459 37.828 114.796 1.00 69.14 0 ATOM 722 CD1 TYR78 11.822 36.717 115.368 1.00 76.95 0 ATOM 723 CE1 TYR 78 12.278 36.142116.548 1.00 77.96 0 ATOM 724 CD2 TYR 78 13.577 38.346 115.446 1.0075.81 0 ATOM 725 CE2 TYR 78 14.044 37.775 116.638 1.00 77.77 0 ATOM 726CZ TYR 78 13.387 36.673 117.181 1.00 77.18 0 ATOM 727 OH TYR 78 13.82636.115 118.365 1.00 87.14 0 ATOM 729 C TYR 78 9.967 36.964 113.053 1.0015.29 0 ATOM 730 O TYR 78 9.811 35.796 113.403 1.00 62.78 0 ATOM 731 NGLY 79 8.992 37.863 113.117 1.00 2.00 0 ATOM 733 CA GLY 79 7.675 37.485113.582 1.00 2.00 0 ATOM 734 C GLY 79 6.977 36.532 112.629 1.00 2.00 0ATOM 735 O GLY 79 6.226 35.649 113.052 1.00 34.11 0 ATOM 736 N GLY 807.237 36.699 111.338 1.00 49.23 0 ATOM 738 CA GLY 80 6.601 35.870110.330 1.00 48.34 0 ATOM 739 C GLY 80 5.699 36.782 109.520 1.00 47.91 0ATOM 740 O GLY 80 4.955 37.572 110.097 1.00 15.83 0 ATOM 741 N PHE 815.759 36.692 108.193 1.00 2.00 0 ATOM 743 CA PHE 81 4.948 37.558 107.3431.00 2.00 0 ATOM 744 CB PHE 81 5.386 37.403 105.880 1.00 2.00 0 ATOM 745CG PHE 81 6.673 38.134 105.556 1.00 2.00 0 ATOM 746 CD1 PHE 81 7.89937.506 105.685 1.00 2.00 0 ATOM 747 CD2 PHE 81 6.652 39.467 105.139 1.002.00 0 ATOM 748 CE1 PHE 81 9.082 38.193 105.405 1.00 2.00 0 ATOM 749 CE2PHE 81 7.829 40.154 104.859 1.00 2.00 0 ATOM 750 CZ PHE 81 9.044 39.517104.992 1.00 2.00 0 ATOM 751 C PHE 81 3.428 37.380 107.548 1.00 2.00 0ATOM 752 O PHE 81 2.918 36.257 107.636 1.00 2.00 0 ATOM 753 N PRO 822.694 38.505 107.618 1.00 2.00 0 ATOM 754 CD PRO 82 3.317 39.807 107.3371.00 21.23 0 ATOM 755 CA PRO 82 1.261 38.702 107.820 1.00 2.00 0 ATOM756 CB PRO 82 0.996 39.979 107.079 1.00 20.78 0 ATOM 757 CG PRO 82 2.14440.765 107.502 1.00 22.98 0 ATOM 758 C PRO 82 0.273 37.623 107.490 1.002.00 0 ATOM 759 O PRO 82 −0.761 37.529 108.161 1.00 36.87 0 ATOM 760 NPRO 83 0.501 36.841 106.423 1.00 27.66 0 ATOM 761 CD PRO 83 1.439 36.880105.290 1.00 2.85 0 ATOM 762 CA PRO 83 −0.514 35.812 106.201 1.00 30.550 ATOM 763 CB PRO 83 0.089 34.958 105.083 1.00 5.44 0 ATOM 764 CG PRO 831.550 35.427 104.971 1.00 10.08 0 ATOM 765 C PRO 83 −0.791 34.990107.460 1.00 24.53 0 ATOM 766 O PRO 83 −1.947 34.863 107.882 1.00 5.06 0ATOM 767 N GLU 84 0.284 34.504 108.080 1.00 63.06 0 ATOM 769 CA GLU 840.190 33.665 109.268 1.00 65.66 0 ATOM 770 CB GLU 84 1.448 32.797109.382 1.00 61.87 0 ATOM 771 CG GLU 84 1.194 31.402 109.979 1.00 75.070 ATOM 772 CD GLU 84 0.401 30.452 109.056 1.00 79.88 0 ATOM 773 OE1 GLU84 1.015 29.508 108.504 1.00 76.25 0 ATOM 774 OE2 GLU 84 −0.832 30.635108.891 1.00 74.31 0 ATOM 775 C GLU 84 −0.066 34.394 110.588 1.00 66.470 ATOM 776 O GLU 84 −1.194 34.406 111.085 1.00 61.22 0 ATOM 777 N SER 850.984 34.978 111.161 1.00 66.52 0 ATOM 779 CA SER 85 0.882 35.691112.431 1.00 64.76 0 ATOM 780 CB SER 85 2.266 36.145 112.900 1.00 2.00 0ATOM 781 OG SER 85 3.198 35.078 112.871 1.00 2.00 0 ATOM 783 C SER 85−0.013 36.917 112.349 1.00 62.83 0 ATOM 784 O SER 85 −0.005 37.633111.341 1.00 2.00 0 ATOM 785 N ASN 86 −0.785 37.146 113.412 1.00 2.00 0ATOM 787 CA ASN 86 −1.647 38.321 113.490 1.00 2.00 0 ATOM 788 CB ASN 86−2.747 38.123 114.520 1.00 2.00 0 ATOM 789 CG ASN 86 −3.887 37.283113.994 1.00 10.84 0 ATOM 790 OD1 ASN 86 −3.763 36.629 112.957 1.0012.15 0 ATOM 791 ND2 ASN 86 −5.017 37.300 114.705 1.00 6.45 0 ATOM 794 CASN 86 −0.742 39.467 113.912 1.00 2.00 0 ATOM 795 O ASN 86 0.363 39.229114.423 1.00 2.00 0 ATOM 796 N TYR 87 −1.183 40.703 113.723 1.00 2.00 0ATOM 798 CA TYR 87 −0.337 41.830 114.081 1.00 2.00 0 ATOM 799 CB TYR 870.529 42.253 112.870 1.00 2.00 0 ATOM 800 CG TYR 87 1.802 41.448 112.6841.00 2.00 0 ATOM 801 CD1 TYR 87 1.879 40.421 111.749 1.00 2.00 0 ATOM802 CE1 TYR 87 3.039 39.689 111.598 1.00 2.00 0 ATOM 803 CD2 TYR 872.930 41.714 113.458 1.00 2.00 0 ATOM 804 CE2 TYR 87 4.080 40.991113.313 1.00 2.00 0 ATOM 805 CZ TYR 87 4.127 39.986 112.384 1.00 2.00 0ATOM 806 OH TYR 87 5.280 39.281 112.259 1.00 2.00 0 ATOM 808 C TYR 87−1.104 43.029 114.588 1.00 2.00 0 ATOM 809 O TYR 87 −2.264 43.238114.216 1.00 2.00 0 ATOM 810 N LEU 88 −0.440 43.810 115.435 1.00 2.00 0ATOM 812 CA LEU 88 −1.015 45.026 115.987 1.00 2.00 0 ATOM 813 CB LEU 88−1.615 44.766 117.375 1.00 2.00 0 ATOM 814 CG LEU 88 −2.364 45.917118.062 1.00 2.00 0 ATOM 815 CD1 LEU 88 −3.657 46.225 117.348 1.00 2.000 ATOM 816 CD2 LEU 88 −2.675 45.551 119.493 1.00 2.00 0 ATOM 817 C LEU88 0.119 46.034 116.084 1.00 2.00 0 ATOM 818 O LEU 88 1.082 45.814116.814 1.00 2.00 0 ATDM 819 N PHE 89 0.061 47.108 115.309 1.00 2.00 0ATOM 821 CA PHE 89 1.114 48.117 115.393 1.00 2.00 0 ATOM 822 CB PHE 891.567 48.599 114.004 1.00 2.00 0 ATOM 823 CG PHE 89 2.305 47.563 113.2211.00 2.00 0 ATOM 824 CD1 PHE 89 1.617 46.543 112.578 1.00 2.00 0 ATOM825 CD2 PHE 89 3.683 47.581 113.159 1.00 2.00 0 ATOM 826 CE1 PHE 892.284 45.547 111.885 1.00 2.00 0 ATOM 827 CE2 PHE 89 4.379 46.586112.464 1.00 2.00 0 ATOM 828 CZ PHE 89 3.673 45.565 111.826 1.00 2.00 0ATOM 829 C PHE 89 0.495 49.250 116.197 1.00 2.00 0 ATOM 830 O PHE 89−0.664 49.615 115.985 1.00 2.00 0 ATOM 831 N LEU 90 1.271 49.797 117.1241.00 4.99 0 ATOM 833 CA LEU 90 0.810 50.856 118.014 1.00 4.99 0 ATOM 834CB LEU 90 1.519 50.702 119.370 1.00 2.00 0 ATOM 835 CG LEU 90 1.51649.291 119.991 1.00 2.00 0 ATOM 836 CD1 LEU 90 2.488 49.246 121.147 1.002.00 0 ATOM 837 CD2 LEU 90 0.111 48.870 120.402 1.00 2.00 0 ATOM 838 CLEU 90 1.000 52.275 117.463 1.00 4.99 0 ATOM 839 O LEU 90 1.382 53.196118.200 1.00 2.00 0 ATOM 840 N GLY 91 0.734 52.459 116.175 1.00 14.03 0ATOM 842 CA GLY 91 0.884 53.774 115.588 1.00 14.03 0 ATOM 843 C GLY 912.307 54.139 115.209 1.00 14.03 0 ATOM 844 O GLY 91 3.220 53.312 115.2671.00 32.27 0 ATOM 845 N ASP 92 2.471 55.400 114.820 1.00 2.00 0 ATOM 847CA ASP 92 3.736 55.978 114.382 1.00 2.00 0 ATOM 848 CB ASP 92 4.67056.170 115.576 1.00 2.00 0 ATOM 849 CG ASP 92 4.185 57.239 116.525 1.002.00 0 ATOM 850 OD1 ASP 92 3.449 56.145 116.063 1.00 2.00 0 ATOM 851 OD2ASP 92 4.541 57.177 117.725 1.00 2.00 0 ATOM 852 C ASP 92 4.427 55.205113.259 1.00 2.00 0 ATOM 853 O ASP 92 5.515 54.631 113.422 1.00 2.00 0ATOM 854 N TYR 93 3.792 55.234 112.096 1.00 2.00 0 ATOM 856 CA TYR 934.301 54.523 110.938 1.00 2.00 0 ATOM 857 CB TYR 93 3.149 53.907 110.1711.00 2.00 0 ATOM 858 CG TYR 93 2.122 53.226 111.029 1.00 2.00 0 ATOM 859CD1 TYR 93 0.875 53.801 111.227 1.00 2.00 0 ATOM 860 CE1 TYR 93 −0.08653.176 112.004 1.00 2.00 0 ATOM 861 CD2 TYR 93 2.390 52.002 111.628 1.002.00 0 ATOM 862 CE2 TYR 93 1.445 51.362 112.405 1.00 2.00 0 ATOM 863 CZTYR 93 0.204 51.951 112.593 1.00 2.00 0 ATOM 864 OH TYR 93 −0.736 51.315113.379 1.00 2.00 0 ATOM 866 C TYR 93 5.088 55.399 109.992 1.00 2.00 0ATOM 867 O TYR 93 5.998 54.922 109.335 1.00 2.00 0 ATOM 868 N VAL 944.718 56.667 109.904 1.00 2.00 0 ATOM 870 CA VAL 94 5.380 57.607 109.0041.00 2.00 0 ATOM 871 CB VAL 94 4.322 58.340 108.112 1.00 2.00 0 ATOM 872CG1 VAL 94 3.365 57.315 107.523 1.00 2.00 0 ATOM 873 CG2 VAL 94 3.55159.356 108.903 1.00 2.00 0 ATOM 874 C VAL 94 6.253 58.614 109.780 1.002.00 0 ATOM 875 O VAL 94 6.447 58.457 110.985 1.00 2.00 0 ATOM 876 N ASP95 6.774 59.629 109.094 1.00 8.40 0 ATOM 878 CA ASP 95 7.641 60.648109.689 1.00 2.00 0 ATOM 879 CB ASP 95 6.967 61.356 110.868 1.00 39.32 0ATOM 880 CG ASP 95 5.975 62.433 110.449 1.00 45.35 0 ATOM 881 OD1 ASP 956.227 63.167 109.471 1.00 44.75 0 ATOM 882 OD2 ASP 95 4.937 62.561111.127 1.00 53.79 0 ATOM 883 C ASP 95 8.975 60.074 110.155 1.00 2.71 0ATOM 884 O ASP 95 9.092 58.877 110.421 1.00 37.69 0 ATOM 885 N ARG 969.972 60.949 110.252 1.00 21.97 0 ATOM 887 CA ARG 96 11.322 60.589110.685 1.00 27.39 0 ATOM 888 CB ARG 96 11.285 59.989 112.099 1.00 18.630 ATOM 889 CG ARG 96 12.037 60.816 113.128 1.00 24.81 0 ATOM 890 CD ARG96 11.255 62.052 113.581 1.00 32.64 0 ATOM 891 NE ARG 96 10.551 61.833114.850 1.00 39.46 0 ATOM 893 CZ ARG 96 9.890 62.774 115.525 1.00 39.860 ATOM 894 NH1 ARG 96 9.824 64.018 115.066 1.00 40.95 0 ATOM 897 NH2 ARG96 9.290 62.471 116.670 1.00 44.56 0 ATOM 900 C ARG 96 12.109 59.659109.732 1.00 24.24 0 ATOM 901 C ARG 96 13.114 60.072 109.135 1.00 13.090 ATOM 902 N GLY 97 11.668 58.411 109.592 1.00 19.94 0 ATOM 904 CA GLY97 12.359 57.480 108.716 1.00 20.45 0 ATOM 905 C GLY 97 12.412 57.920107.269 1.00 23.76 0 ATOM 906 O GLY 97 11.516 58.617 106.773 1.00 84.640 ATOM 907 N LYS 98 13.460 57.469 106.584 1.00 47.57 0 ATOM 909 CA LYS98 13.698 57.806 105.182 1.00 46.79 0 ATOM 910 CB LYS 98 15.147 57.462104.832 1.00 31.45 0 ATOM 911 CG LYS 98 16.169 58.397 105.487 1.00 30.730 ATOM 912 CD LYS 98 17.606 59.061 105.087 1.00 32.79 0 ATOM 913 CE LYS98 18.605 59.089 105.627 1.00 30.08 0 ATOM 914 NZ LYS 98 20.036 58.743105.315 1.00 31.54 0 ATOM 918 C LYS 98 12.741 57.196 104.146 1.00 44.780 ATOM 919 O LYS 98 12.613 57.707 103.040 1.00 31.43 0 ATOM 920 N GLN 9912.059 56.120 104.517 1.00 2.00 0 ATOM 922 CA GLN 99 11.132 55.430103.639 1.00 2.00 0 ATOM 923 CB GLN 99 11.654 54.023 103.337 1.00 11.210 ATOM 924 CG GLN 99 12.945 53.993 102.552 1.00 6.66 0 ATOM 925 CD GLN99 13.361 52.586 102.177 1.00 11.01 0 ATOM 926 OE1 GLN 99 13.802 51.805103.031 1.00 10.44 0 ATOM 927 NE2 GLN 99 13.229 52.247 100.895 1.00 8.180 ATOM 930 C GLN 99 9.741 55.328 104.245 1.00 2.00 0 ATOM 931 O GLN 999.177 54.238 104.317 1.00 6.66 0 ATOM 932 N SER 100 9.178 56.460 104.6571.00 24.01 0 ATOM 934 CA SER 100 7.839 56.475 105.257 1.00 24.01 0 ATOM935 CB SER 100 7.481 57.897 105.739 1.00 2.00 0 ATOM 936 OG SER 1008.479 58.490 106.556 1.00 2.00 0 ATOM 938 C SBR 100 6.749 55.987 104.2721.00 24.01 0 ATOM 939 O SER 100 5.703 55.463 104.680 1.00 2.00 0 ATOM940 N LEU 101 7.015 56.153 102.977 1.00 27.83 0 ATOM 942 CA LEU 1016.064 55.786 101.913 1.00 27.83 0 ATOM 943 CB LEU 101 6.551 56.416100.599 1.00 3.66 0 ATOM 944 CG LEU 101 5.593 57.224 99.721 1.00 3.66 0ATOM 945 CD1 LEU 101 4.209 56.591 99.749 1.00 3.66 0 ATOM 946 CD2 LEU101 5.532 58.653 100.208 1.00 3.66 0 ATOM 947 C LEU 101 5.852 54.284101.689 1.00 27.83 0 ATOM 948 O LEU 101 4.731 53.851 101.467 1.00 3.66 0ATOM 949 N GLU 102 6.901 53.480 101.725 1.00 2.00 0 ATOM 951 CA GLU 1026.713 52.055 101.490 1.00 2.00 0 ATOM 952 CB GLU 102 7.976 51.470100.870 1.00 13.98 0 ATOM 953 CG GLU 102 9.211 52.208 101.294 1.00 13.980 ATOM 954 CD GLU 102 10.116 52.539 100.136 1.00 13.98 0 ATOM 955 OE1GLU 102 10.151 53.727 99.737 1.00 13.98 0 ATOM 956 OE2 GLU 102 10.77251.599 99.643 1.00 13.98 0 ATOM 957 C GLU 102 6.307 51.324 102.763 1.002.00 0 ATOM 958 O GLU 102 5.686 50.263 102.729 1.00 13.98 0 ATOM 959 NTHR 103 6.664 51.901 103.897 1.00 2.00 0 ATOM 961 CA THR 103 6.29351.330 105.173 1.00 2.00 0 ATOM 962 CB THR 103 6.923 52.098 106.309 1.002.00 0 ATOM 963 OG1 THR 103 8.313 52.297 106.028 1.00 2.00 0 ATOM 965CG2 THR 103 6.758 51.329 107.599 1.00 2.00 0 ATOM 966 C THR 103 4.77551.440 105.296 1.00 2.00 0 ATOM 967 O THR 103 4.081 50.428 105.394 1.002.00 0 ATOM 968 N ILE 104 4.256 52.667 105.257 1.00 2.00 0 ATOM 970 CAILE 104 2.824 52.873 105.363 1.00 2.00 0 ATOM 971 CB ILE 104 2.48654.389 105.300 1.00 12.72 0 ATOM 972 CG2 ILE 104 2.856 54.965 103.9561.00 12.35 0 ATOM 973 CG1 ILE 104 0.996 54.615 105.548 1.00 13.84 0 ATOM974 CD1 ILE 104 0.459 53.940 106.787 1.00 12.35 0 ATOM 975 C ILE 1042.074 52.062 104.294 1.00 2.00 0 ATOM 976 O ILE 104 0.975 51.581 104.5441.00 18.28 0 ATOM 977 N CYS 105 2.690 51.868 103.127 1.00 23.45 0 ATOM979 CA CYS 105 2.080 51.092 102.042 1.00 21.74 0 ATOM 980 CB CYS 1052.722 51.458 100.708 1.00 20.76 0 ATOM 981 SG CYS 105 2.010 52.95899.957 1.00 18.03 0 ATOM 982 C CYS 105 2.061 49.563 102.214 1.00 17.08 0ATOM 983 O CYS 105 1.114 48.915 101.781 1.00 20.76 0 ATOM 984 N LEU 1063.089 48.952 102.833 1.00 2.00 0 ATOM 986 CA LEU 106 3.124 47.526103.073 1.00 2.00 0 ATOM 987 CB LEU 106 4.519 47.042 103.495 1.00 2.00 0ATOM 988 CG LEU 106 4.680 45.544 103.802 1.00 2.00 0 ATOM 989 CD1 LEU106 4.200 44.694 102.639 1.00 2.00 0 ATOM 990 CD2 LEU 106 6.133 45.247104.053 1.00 2.00 0 ATOM 991 C LEU 106 2.154 47.179 104.178 1.00 2.00 0ATOM 992 O LEU 106 1.589 46.088 104.192 1.00 2.00 0 ATOM 993 N LEU 1071.992 48.107 105.116 1.00 2.00 0 ATOM 995 CA LEU 107 1.078 47.922106.226 1.00 2.00 0 ATOM 996 CB LEU 107 1.347 48.967 107.319 1.00 2.00 0ATOM 997 CG LEU 107 2.761 48.857 107.911 1.00 2.00 0 ATOM 998 CD1 LEU107 3.106 50.016 108.815 1.00 2.00 0 ATOM 999 CD2 LEU 107 2.868 47.535108.638 1.00 2.00 0 ATOM 1000 C LEU 107 −0.339 48.035 105.683 1.00 2.000 ATOM 1001 O LEU 107 −1.153 47.125 105.873 1.00 2.00 0 ATOM 1002 N LEU108 −0.623 49.126 104.971 1.00 2.00 0 ATOM 1004 CA LEU 108 −1.953 49.340104.394 1.00 2.00 0 ATOM 1005 CB LEU 108 −2.020 50.666 103.637 1.00 2.000 ATOM 1006 CG LEU 108 −2.103 51.925 104.499 1.00 2.00 0 ATOM 1007 CD1LEU 108 −2.244 53.164 103.618 1.00 2.00 0 ATOM 1008 CD2 LEU 108 −3.28151.792 105.433 1.00 2.00 0 ATOM 1009 C LEU 108 −2.352 48.206 103.4551.00 2.00 0 ATOM 1010 O LEU 108 −3.533 47.846 103.369 1.00 2.00 0 ATOM1011 N ALA 109 −1.366 47.643 102.762 1.00 2.00 0 ATOM 1013 CA ALA 109−1.608 46.539 101.839 1.00 2.00 0 ATOM 1014 CB ALA 109 −0.336 46.209101.087 1.00 33.06 0 ATOM 1015 C ALA 109 −2.091 45.316 102.605 1.00 2.000 ATOM 1016 O ALA 109 −3.146 44.746 102.305 1.00 24.92 0 ATOM 1017 N TYR110 −1.311 44.929 103.609 1.00 5.11 0 ATOM 1019 CA TYR 110 −1.625 43.781104.452 1.00 2.00 0 ATOM 1020 CB TYR 110 −0.495 43.564 105.438 1.00 2.000 ATOM 1021 CG TYR 110 0.674 42.771 104.922 1.00 2.00 0 ATOM 1022 CD1TYR 110 1.966 43.235 105.092 1.00 2.00 0 ATOM 1023 CE1 TYR 110 3.05342.474 104.706 1.00 2.00 0 ATOM 1024 CD2 TYR 110 0.494 41.517 104.3401.00 2.00 0 ATOM 1025 CE2 TYR 110 1.578 40.743 103.950 1.00 2.00 0 ATOM1026 CZ TYR 110 2.859 41.230 104.139 1.00 2.00 0 ATOM 1027 OH TYR 1103.963 40.483 103.779 1.00 2.00 0 ATOM 1029 C TYR 110 −2.948 43.940105.215 1.00 2.00 0 ATOM 1030 O TYR 110 −3.663 42.961 105.441 1.00 2.000 ATOM 1031 N LYS 111 −3.265 45.168 105.618 1.00 2.00 0 AYOM 1033 CA LYS111 −4.508 45.430 106.333 1.00 2.00 0 ATOM 1034 CB LYS 111 −4.619 46.904106.708 1.00 2.00 0 ATOM 1035 CG LYS 111 −5.942 47.262 107.393 1.00 2.000 ATOM 1036 CD LYS 111 −6.065 46.501 108.685 1.00 2.00 0 ATOM 1037 CELYS 111 −7.410 46.743 109.354 1.00 2.00 0 ATOM 1038 NZ LYS 111 −7.64345.735 110.428 1.00 2.00 0 ATOM 1042 C LYS 111 −5.698 45.051 105.4651.00 2.00 0 ATOM 1043 O LYS 111 −6.655 44.431 105.951 1.00 2.00 0 ATOM1044 N ILE 112 −5.624 45.455 104.192 1.00 2.00 0 ATOM 1046 CA ILE 112−6.651 45.192 103.177 1.00 2.00 0 ATOM 1047 CB ILE 112 −6.361 45.978101.875 1.00 25.04 0 ATOM 1048 CG2 ILE 112 −7.414 45.656 100.814 1.0025.04 0 ATOM 1049 CG1 ILE 112 −6.339 47.482 102.166 1.00 25.04 0 ATOM1050 CD1 ILE 112 −5.857 48.336 101.012 1.00 25.04 0 ATOM 1051 C ILE 112−6.706 43.706 102.829 1.00 2.00 0 ATOM 1052 O ILE 112 −7.793 43.156102.568 1.00 25.04 0 ATOM 1053 N LYS 113 −5.537 43.067 102.816 1.00 2.000 ATOM 1055 CA LYS 113 5.447 41.649 102.513 1.00 2.00 0 ATOM 1056 CB LYS113 −4.001 41.285 102.181 1.00 8.72 0 ATOM 1057 CG LYS 113 −3.852 39.909101.596 1.00 8.72 0 ATOM 1058 CD LYS 113 −2.780 39.871 100.521 1.00 8.720 ATOM 1059 CE LYS 113 −2.618 38.467 99.975 1.00 8.72 0 ATOM 1060 NZ LYS113 −3.952 37.873 99.637 1.00 8.72 0 ATOM 1064 C LYS 113 −5.987 40.780103.667 1.00 2.00 0 ATOM 1065 O LYS 113 −6.620 39.742 103.435 1.00 8.720 ATOM 1066 N TYR 114 −5.744 41.200 104.906 1.00 9.02 0 ATOM 1068 CA TYR114 −6.226 40.455 106.068 1.00 8.11 0 ATOM 1069 CB TYR 114 −5.122 39.580106.666 1.00 12.47 0 ATOM 1070 CC TYR 114 −4.138 38.988 105.689 1.0012.47 0 ATOM 1071 CD1 TYR 114 −3.027 39.715 105.278 1.00 12.47 0 ATOM1072 CE1 TYR 114 −2.111 39.191 104.397 1.00 12.47 0 ATOM 1073 CD2 TYR114 −4.307 37.709 105.188 1.00 12.47 0 ATOM 1074 CE2 TYR 114 −3.39537.168 104.302 1.00 12.47 0 ATOM 1075 CZ TYR 114 −2.296 37.917 103.9081.00 12.47 0 ATOM 1076 OH TYR 114 −1.378 37.404 103.013 1.00 12.47 0ATOM 1078 C TYR 114 −6.729 41.417 107.155 1.00 9.76 0 ATOM 1079 O TYR114 −6.058 41.636 108.169 1.00 12.47 0 ATOM 1080 N PRO 115 −7.928 41.985106.968 1.00 34.30 0 ATOM 1081 CD PRO 115 −8.843 41.845 105.826 1.0024.86 0 ATOM 1082 CA PRO 115 −8.488 42.912 107.944 1.00 34.30 0 ATOM1083 CB PRO 115 −9.862 43.227 107.365 1.00 24.86 0 ATOM 1084 CG PRO 115−9.640 43.117 105.921 1.00 24.86 0 ATOM 1085 C PRO 115 −8.586 42.310109.335 1.00 34.30 0 ATOM 1086 O PRO 115 −8.017 42.937 110.285 1.0024.86 0 ATOM 1087 N GLU 116 −9.275 41.181 109.445 1.00 2.00 0 ATOM 1089CA GLU 116 −9.477 40.538 110.742 1.00 2.00 0 ATOM 1090 CB GLU 116−10.577 39.469 110.637 1.00 35.30 0 ATOM 1091 CG GLU 116 −11.673 39.726109.593 1.00 37.08 0 ATOM 1092 CD GLU 116 −12.739 40.728 110.027 1.0043.38 0 ATOM 1093 OE1 GLU 116 −13.060 41.641 109.235 1.00 47.93 0 ATOM1094 OE2 GLU 116 −13.274 40.599 111.147 1.00 48.80 0 ATOM 1095 C GLU 116−8.213 39.902 111.358 1.00 2.00 0 ATOM 1096 O GLU 116 −8.296 39.287112.424 1.00 32.95 0 ATOM 1097 N ASN 117 −7.054 40.067 110.712 1.00 2.000 ATOM 1099 CA ASN 117 −5.820 39.455 111.211 1.00 2.00 0 ATOM 1100 CBASN 117 −5.457 38.220 110.375 1.00 6.12 0 ATOM 1101 CG ASN 117 −6.55237.174 110.353 1.00 8.10 0 ATOM 1102 OD1 ASN 117 −7.584 37.351 109.7001.00 17.57 0 ATOM 1103 ND2 ASN 117 −6.328 36.071 111.048 1.00 14.33 0ATOM 1106 C ASN 117 −4.613 40.372 111.211 1.00 2.00 0 ATOM 1107 O ASN117 −3.496 39.952 111.506 1.00 8.94 0 ATOM 1108 N PHE 118 −4.821 41.620110.862 1.00 2.00 0 ATOM 1110 CA PHE 118 −3.715 42.559 110.815 1.00 2.000 ATOM 1111 CB PHE 118 −3.135 42.617 109.395 1.00 2.00 0 ATOM 1112 CGPHE 118 −1.902 43.471 109.256 1.00 2.00 0 ATOM 1113 CD1 PHE 118 −0.64742.886 109.155 1.00 2.00 0 ATOM 1114 CD2 PHE 118 −1.995 44.857 109.1771.00 2.00 0 ATOM 1115 CE1 PHE 118 0.503 43.680 108.972 1.00 2.00 0 ATOM1116 CE2 PHE 118 −0.657 45.647 108.997 1.00 2.00 0 ATOM 1117 CZ PHE 1180.393 45.060 108.894 1.00 2.00 0 ATOM 1118 C PHE 118 −4.392 43.854111.194 1.00 2.00 0 ATOM 1119 O PHE 118 −5.384 44.259 110.576 1.00 2.000 ATOM 1120 N PHE 119 −3.874 44.483 112.240 1.00 12.11 0 ATOM 1122 CAPHE 119 −4.467 45.700 112.721 1.00 12.11 0 ATOM 1123 CB PHE 119 −5.17445.400 114.022 1.00 2.00 0 ATOM 1124 CG PHE 119 −6.229 44.355 113.8891.00 2.00 0 ATOM 1125 CD1 PHE 119 −5.903 43.008 113.975 1.00 2.00 0 ATOM1126 CD2 PHE 119 −7.556 44.716 113.655 1.00 2.00 0 ATOM 1127 CE1 PHE 119−6.881 42.035 113.829 1.00 2.00 0 ATOM 1128 CE2 PHE 119 −8.546 43.753113.506 1.00 2.00 0 ATOM 1129 CZ PHE 119 −8.209 42.410 113.592 1.00 2.000 ATOM 1130 C PHE 119 −3.472 46.806 112.905 1.00 12.11 0 ATOM 1131 O PHE119 −2.342 46.577 113.346 1.00 2.00 0 ATOM 1132 N LEU 120 −3.893 48.008112.537 1.00 2.00 0 ATOM 1134 CA LEU 120 −3.043 49.179 112.672 1.00 2.000 ATOM 1135 CB LEU 120 −2.770 49.809 111.303 1.00 2.00 0 ATOM 1136 CGLEU 120 −2.127 48.910 110.259 1.00 2.00 0 ATOM 1137 CD1 LEU 120 −2.14749.623 108.924 1.00 2.00 0 ATOM 1138 CD2 LEU 120 −0.731 46.539 110.6851.00 2.00 0 ATOM 1139 C LEU 120 −3.766 50.176 113.559 1.00 2.00 0 ATOM1140 O LEU 120 −4.963 50.401 113.375 1.00 2.00 0 ATOM 1141 N LEU 121−3.069 50.730 114.542 1.00 2.00 0 ATOM 1143 CA LEU 121 −3.670 51.714115.415 1.00 2.00 0 ATOM 1144 CB LEU 121 −3.351 51.428 116.890 1.00 2.000 ATOM 1145 CG LEU 121 −4.142 50.320 117.598 1.00 2.00 0 ATOM 1146 CD1LEU 121 −3.648 50.150 119.012 1.00 2.00 0 ATOM 1147 CD2 LEU 121 −5.60950.657 117.581 1.00 2.00 0 ATOM 1148 C LEU 121 −3.100 53.060 115.0041.00 2.00 0 ATOM 1149 O LEU 121 −2.213 53.148 114.166 1.00 2.00 0 ATOM1150 N ARG 122 −3.631 54.118 115.592 1.00 2.00 0 ATOM 1152 CA ARG 122−3.162 55.434 115.251 1.00 2.00 0 ATOM 1153 CB ARG 122 −4.336 56.404115.224 1.00 2.00 0 ATOM 1154 CG ARG 122 −4.047 57.765 114.619 1.00 2.000 ATOM 1155 CD ARG 122 −5.298 58.565 114.683 1.00 2.00 0 ATOM 1156 NEARG 122 −5.207 59.851 114.014 1.00 2.00 0 ATOM 1158 CZ ARG 122 −6.27460.578 113.685 1.00 2.00 0 ATOM 1159 NH1 ARG 122 −7.512 60.131 113.9561.00 2.00 0 ATOM 1162 NH2 ARG 122 −6.104 61.762 113.102 1.00 2.00 0 ATOM1165 C ARG 122 −2.104 55.942 116.208 1.00 2.00 0 ATOM 1166 O ARG 122−2.163 55.716 117.423 1.00 2.00 0 ATOM 1167 N GLY 123 −1.135 56.635115.631 1.00 2.00 0 ATOM 1169 CA GLY 123 −0.073 57.235 116.399 1.00 2.000 ATOM 1170 C GLY 123 −0.205 58.730 116.198 1.00 2.00 0 ATOM 1171 O GLY123 −0.865 59.177 115.262 1.00 86.19 0 ATOM 1172 N ASN 124 0.437 59.508117.058 1.00 2.00 0 ATOM 1174 CA ASN 124 0.390 60.956 116.958 1.00 2.000 ATOM 1175 CB ASN 124 1.003 61.584 118.199 1.00 4.11 0 ATOM 1176 CG ASN124 2.477 61.333 118.313 1.00 8.88 0 ATOM 1177 OD1 ASN 124 2.940 60.187118.486 1.00 8.54 0 ATOM 1178 ND2 ASN 124 3.240 62.400 118.236 1.00 6.380 ATOM 1181 C ASN 124 1.103 61.455 115.708 1.00 2.00 0 ATOM 1182 O ASN124 1.143 62.664 115.430 1.00 5.53 0 ATOM 1183 N HIS 125 1.678 60.515114.963 1.00 18.81 0 ATOM 1185 CA HIS 125 2.372 60.815 113.723 1.0019.06 0 ATOM 1186 CB HIS 125 3.759 60.186 113.744 1.00 13.41 0 ATOM 1187CG HIS 125 4.790 61.072 114.362 1.00 13.65 0 ATOM 1188 CD2 HIS 125 4.76561.811 115.493 1.00 8.85 0 ATOM 1189 ND1 HIS 125 6.006 61.322 113.7721.00 14.49 0 ATOM 1191 CE1 HIS 125 6.686 62.182 114.503 1.00 13.93 0ATOM 1192 NE2 HIS 125 5.954 62.496 115.554 1.00 12.46 0 ATOM 1194 C HIS125 1.555 60.338 112.523 1.00 16.39 0 ATOM 1195 O HIS 125 2.090 59.905111.513 1.00 13.34 0 ATOM 1196 N GLU 126 0.241 60.382 112.686 1.00 2.000 ATOM 1198 CA GLU 126 −0.731 60.018 111.664 1.00 2.00 0 ATOM 1199 CBGLU 126 −1.462 58.725 112.050 1.00 2.00 0 ATOM 1200 CG GLU 126 −0.78357.443 111.570 1.00 2.00 0 ATOM 1201 CD GLU 126 0.578 57.217 112.1901.00 2.00 0 ATOM 1202 OE1 GLU 126 1.615 57.346 111.504 1.00 2.00 0 ATOM1203 OE2 GLU 126 0.610 56.898 113.385 1.00 2.00 0 ATOM 1204 C GLU 126−1.683 61.211 111.705 1.00 2.00 0 ATOM 1205 O GLU 126 −2.903 61.073111.837 1.00 2.00 0 ATOM 1206 N CYS 127 −1.097 62.392 111.578 1.00 2.000 ATOM 1208 CA CYS 127 −1.860 63.608 111.681 1.00 2.00 0 ATOM 1209 CBCYS 127 −2.037 63.960 113.167 1.00 26.08 0 ATOM 1210 SG CYS 127 −3.05265.420 113.546 1.00 39.49 0 ATOM 1211 C CYS 127 −1.142 64.731 110.9761.00 2.00 0 ATOM 1212 O CYS 127 0.048 64.974 111.204 1.00 23.46 0 ATOM1213 N ALA 128 −1.912 65.421 110.142 1.00 2.00 0 ATOM 1215 CA ALA 128−1.489 66.551 109.335 1.00 2.00 0 ATOM 1216 CB ALA 128 −2.686 67.412109.031 1.00 2.00 0 ATOM 1217 C ALA 128 −0.385 67.413 109.910 1.00 2.000 ATOM 1218 O ALA 128 0.690 67.456 109.346 1.00 2.00 0 ATOM 1219 N SER129 −0.649 68.097 111.021 1.00 2.00 0 ATOM 1221 CA SER 129 0.331 68.981111.675 1.00 2.00 0 ATOM 1222 CB SER 129 −0.288 69.580 112.937 1.0028.29 0 ATOM 1223 OG SER 129 −0.836 68.562 113.760 1.00 32.64 0 ATOM1225 C SER 129 1.671 68.344 112.042 1.00 2.00 0 ATOM 1226 O SER 1292.669 69.048 112.186 1.00 24.43 0 ATOM 1227 N ILE 130 1.687 67.027112.226 1.00 2.00 0 ATOM 1229 CA ILE 130 2.920 66.343 112.572 1.00 2.000 ATOM 1230 CB ILE 130 2.671 65.177 113.588 1.00 2.00 0 ATOM 1231 CG2ILE 130 3.999 64.584 114.058 1.00 2.00 0 ATOM 1232 CG1 ILE 130 2.01265.717 114.854 1.00 2.00 0 ATOM 1233 CD1 ILE 130 2.828 66.820 115.5541.00 2.00 0 ATOM 1234 C ILE 130 3.563 65.829 111.289 1.00 2.00 0 ATOM1235 O ILE 130 4.776 65.962 111.113 1.00 2.00 0 ATOM 1236 N ASN 1312.751 65.253 110.397 1.00 15.50 0 ATOM 1238 CA ASN 131 3.209 64.747109.091 1.00 12.35 0 ATOM 1239 CB ASN 131 2.027 64.290 108.231 1.00 2.000 ATOM 1240 CG ASN 131 1.487 62.929 108.632 1.00 2.00 0 ATOM 1241 OD1ASN 131 1.712 62.438 109.750 1.00 2.00 0 ATOM 1242 ND2 ASN 131 0.74562.313 107.718 1.00 2.00 0 ATOM 1245 C ASN 131 3.849 65.914 106.374 1.0028.24 0 ATOM 1246 O ASN 131 5.001 65.854 107.951 1.00 2.00 0 ATOM 1247 NARG 132 3.051 66.971 108.255 1.00 2.00 0 ATOM 1249 CA ARG 132 3.38666.234 107.627 1.00 2.00 0 ATOM 1250 CB ARG 132 2.327 69.269 108.0061.00 22.25 0 ATOM 1251 CG ARG 132 2.505 70.673 107.477 1.00 26.48 0 ATOM1252 CD ARG 132 2.505 70.734 105.962 1.00 27.28 0 ATOM 1253 NE ARG 1323.849 70.533 105.434 1.00 36.02 0 ATOM 1255 CZ ARG 132 4.577 71.479104.852 1.00 34.03 0 ATOM 1256 NH1 ARG 132 4.080 72.697 104.711 1.0030.29 0 ATOM 1259 NH2 ARG 132 5.808 71.211 104.424 1.00 37.77 0 ATOM1262 C ARG 132 4.768 68.731 107.990 1.00 2.00 0 ATOM 1263 O ARG 1325.359 69.477 107.225 1.00 27.60 0 ATOM 1264 N ILE 133 5.314 68.324109.127 1.00 2.00 0 ATOM 1266 CA ILE 133 6.652 68.601 109.457 1.00 2.000 ATOM 1267 CB ILE 133 6.652 69.644 110.746 1.00 20.12 0 ATOM 1268 CG2ILE 133 6.215 71.055 110.441 1.00 19.34 0 ATOM 1269 CG1 ILE 133 5.76168.999 111.800 1.00 21.76 0 ATOM 1270 CD1 ILE 133 5.687 69.792 113.0651.00 22.74 0 ATOM 1271 C ILE 133 7.794 67.779 109.553 1.00 2.00 0 ATOM1272 O ILE 133 8.915 68.073 109.127 1.00 21.74 0 ATOM 1273 N TYR 1347.521 66.583 110.080 1.00 2.00 0 ATOM 1275 CA TYR 134 8.571 65.567110.270 1.00 2.00 0 ATOM 1276 CB TYR 134 8.330 64.766 111.561 1.00 27.010 ATOM 1277 CG TYR 134 8.270 65.648 112.767 1.00 19.94 0 ATOM 1278 CD1TYR 134 7.092 65.789 113.486 1.00 23.86 0 ATOM 1279 CE1 TYR 134 7.01666.651 114.557 1.00 24.86 0 ATOM 1280 CD2 TYR 134 9.380 66.392 113.1601.00 24.55 0 ATOM 1281 CE2 TYR 134 9.312 67.254 114.228 1.00 23.33 0ATOM 1282 CZ TYR 134 8.128 67.384 114.925 1.00 28.93 0 ATOM 1283 OH TYR134 8.043 68.259 115.984 1.00 27.73 0 ATOM 1285 C TYR 134 8.823 64.606109.139 1.00 2.00 0 ATOM 1286 O TYR 134 8.996 63.404 109.368 1.00 24.620 ATOM 1287 N GLY 135 8.847 65.127 107.922 1.00 11.68 0 ATOM 1289 CA GLY135 9.134 64.275 106.787 1.00 10.40 0 ATOM 1290 C GLY 135 8.028 63.772105.882 1.00 9.03 0 ATOM 1291 O GLY 135 8.010 64.127 104.713 1.00 20.700 ATOM 1292 N PHE 136 7.110 62.963 106.393 1.00 2.00 0 ATOM 1294 CA PHE136 6.062 62.412 105.549 1.00 2.00 0 ATOM 1295 CB PHE 136 4.951 61.757106.347 1.00 2.00 0 ATOM 1296 CG PHE 136 4.091 60.831 105.524 1.00 2.000 ATOM 1297 CD1 PHE 136 4.664 59.734 104.870 1.00 2.00 0 ATOM 1298 CD2PHE 136 2.713 61.036 105.424 1.00 2.00 0 ATOM 1299 CE1 PHE 136 3.88258.846 104.131 1.00 2.00 0 ATOM 1300 CE2 PHE 136 1.915 60.155 104.6881.00 2.00 0 ATOM 1301 CZ PHE 136 2.507 59.050 104.037 1.00 2.00 0 ATOM1302 C PHE 136 5.421 63.358 104.563 1.00 2.00 0 ATOM 1303 O PHE 1365.121 62.926 103.445 1.00 2.00 0 ATOM 1304 N TYR 137 5.199 64.616104.940 1.00 2.00 0 ATOM 1306 CA TYR 137 4.606 65.530 103.977 1.00 2.000 ATOM 1307 CB TYR 137 4.243 66.859 104.598 1.00 2.00 0 ATOM 1308 CG TYR137 3.886 67.921 103.564 1.00 2.00 0 ATOM 1309 CD1 TYR 137 2.550 68.203103.257 1.00 2.00 0 ATOM 1310 CE1 TYR 137 2.228 69.196 102.327 1.00 2.000 ATOM 1311 CD2 TYR 137 4.893 68.666 102.901 1.00 2.00 0 ATOM 1312 CE2TYR 137 4.576 69.644 101.985 1.00 2.00 0 ATOM 1313 CZ TYR 137 3.24769.903 101.707 1.00 2.00 0 ATOM 1314 OH TYR 137 2.920 70.893 100.8231.00 2.00 0 ATOM 1316 C TYR 137 5.566 65.785 102.822 1.00 2.00 0 ATOM1317 O TYR 137 5.187 65.669 101.646 1.00 2.00 0 ATOM 1318 N ASP 1386.802 66.153 103.155 1.00 2.00 0 ATOM 1320 CA ASP 138 7.822 66.439102.147 1.00 2.00 0 ATOM 1321 CB ASP 138 9.102 66.971 102.810 1.00 28.590 ATOM 1322 CG ASP 138 8.832 68.173 103.705 1.00 31.85 0 ATOM 1323 OD1ASP 138 8.999 69.327 103.255 1.00 34.04 0 ATOM 1324 OD2 ASP 138 8.43267.963 104.867 1.00 31.26 0 ATOM 1325 C ASP 138 8.118 65.203 101.3081.00 2.00 0 ATOM 1326 O ASP 138 8.322 65.309 100.102 1.00 27.39 0 ATOM1327 N GLU 139 8.097 64.030 101.926 1.00 31.40 0 ATOM 1329 CA GLU 1398.366 62.797 101.200 1.00 28.32 0 ATOM 1330 C GLU 139 8.380 61.608102.149 1.00 2.00 0 ATOM 1331 CG GLU 139 8.965 60.361 101.526 1.00 4.780 ATOM 1332 CD GLU 139 9.167 59.231 102.525 1.00 3.01 0 ATOM 1333 OE1GLU 139 9.135 58.050 102.087 1.00 2.00 0 ATOM 1334 OE2 GLU 139 9.37259.518 103.738 1.00 4.61 0 ATOM 1335 C GLU 139 7.323 62.572 100.117 1.0030.47 0 ATOM 1336 O GLU 139 7.660 62.419 98.941 1.00 2.00 0 ATOM 1337 NCYS 140 6.055 62.553 100.511 1.00 12.32 0 ATOM 1339 CA CYS 140 4.98162.355 99.553 1.00 3.99 0 ATOM 1340 CB CYS 140 3.625 62.543 100.221 1.0018.56 0 ATOM 1341 SG CYS 140 3.313 61.308 101.477 1.00 23.54 0 ATOM 1342C CYS 140 5.128 63.346 98.416 1.00 12.32 0 ATOM 1343 O CYS 140 5.11762.961 97.257 1.00 21.26 0 ATOM 1344 N LYS 141 5.305 64.616 98.763 1.0041.55 0 ATOM 1346 CA LYS 141 5.446 65.696 97.786 1.00 40.86 0 ATOM 1347CB LYS 141 5.655 67.023 98.531 1.00 38.66 0 ATOM 1348 CG LYS 141 5.85368.246 97.646 1.00 32.19 0 ATOM 1349 CD LYS 141 5.886 69.522 98.468 1.0035.28 0 ATOM 1350 CE LYS 141 5.895 70.750 97.584 1.00 36.09 0 ATOM 1351NZ LYS 141 5.552 71.948 98.380 1.00 40.44 0 ATOM 1355 C LYS 141 6.58665.467 96.791 1.00 40.27 0 ATOM 1356 O LYS 141 6.431 65.642 95.582 1.0028.60 0 ATOM 1357 N ARG 142 7.731 65.073 97.319 1.00 23.84 0 ATOM 1359CA ARG 142 8.912 64.834 96.512 1.00 23.84 0 ATOM 1360 CB ARG 142 10.09764.566 97.444 1.00 22.75 0 ATOM 1361 CG ARG 142 11.368 64.101 96.7781.00 24.10 0 ATOM 1362 CD ARG 142 12.474 64.073 97.799 1.00 33.58 0 ATOM1363 NE ARG 142 12.099 63.283 98.963 1.00 37.62 0 ATOM 1365 CZ ARG 14212.464 62.018 99.150 1.00 46.30 0 ATOM 1366 NH1 ARG 142 13.224 61.40498.249 1.00 44.23 0 ATOM 1369 NH2 ARG 142 12.060 61.363 100.234 1.0042.22 0 ATOM 1372 C ARG 142 8.732 63.689 95.519 1.00 23.84 0 ATOM 1373 OARG 142 8.995 63.838 94.330 1.00 24.93 0 ATOM 1374 N ARG 143 8.26862.550 95.999 1.00 2.00 0 ATOM 1376 CA ARG 143 8.104 61.413 95.125 1.002.00 0 ATOM 1377 CB ARG 143 8.267 60.134 95.941 1.00 2.86 0 ATOM 1378 CGARG 143 9.686 59.941 96.455 1.00 2.86 0 ATOM 1379 CD ARG 143 9.79258.874 97.530 1.00 8.64 0 ATOM 1380 NE ARG 143 9.490 57.527 97.049 1.004.03 0 ATOM 1382 CZ ARG 143 9.492 56.439 97.816 1.00 6.91 0 ATOM 1383NH1 ARG 143 9.782 56.527 99.115 1.00 7.13 0 ATOM 1386 NH2 ARG 143 9.20355.257 97.287 1.00 12.97 0 ATOM 1389 C ARG 143 6.796 61.411 94.367 1.002.00 0 ATOM 1390 O ARG 143 6.707 60.818 93.295 1.00 12.04 0 ATOM 1391 NTYR 144 5.791 62.097 94.897 1.00 2.00 0 ATOM 1393 CA TYR 144 4.45962.120 94.274 1.00 2.00 0 ATOM 1394 CB TYR 144 3.509 61.150 95.021 1.002.00 0 ATOM 1395 CG TYR 144 3.902 59.689 94.953 1.00 2.00 0 ATOM 1396CD1 TYR 144 4.946 59.203 95.716 1.00 2.00 0 ATOM 1397 CE1 TYR 144 5.33657.885 95.635 1.00 2.00 0 ATOM 1398 CD2 TYR 144 3.248 58.804 94.102 1.002.00 0 ATOM 1399 CE2 TYR 144 3.633 57.483 94.017 1.00 2.00 0 ATOM 1400CZ TYR 144 4.683 57.034 94.786 1.00 2.00 0 ATOM 1401 OH TYR 144 5.11355.738 94.703 1.00 2.00 0 ATOM 1403 C TYR 144 3.789 63.502 94.169 1.002.00 0 ATOM 1404 O TYR 144 4.129 64.316 93.297 1.00 2.00 0 ATOM 1405 NASN 145 2.827 63.750 95.058 1.00 2.00 0 ATOM 1407 CA ASN 145 2.08165.000 95.083 1.00 2.00 0 ATOM 1408 CB ASN 145 1.159 65.078 93.867 1.007.57 0 ATOM 1409 CG ASN 145 0.255 63.861 93.748 1.00 5.76 0 ATOM 1410OD1 ASN 145 −0.823 63.826 94.329 1.00 9.42 0 ATOM 1411 ND2 ASN 145 0.70262.851 93.007 1.00 5.88 0 ATOM 1414 C ASN 145 1.229 65.137 96.354 1.002.00 0 ATOM 1415 O ASN 145 0.760 64.143 96.938 1.00 11.61 0 ATOM 1416 NILE 146 1.001 66.391 96.737 1.00 2.00 0 ATOM 1418 CA ILE 146 0.21766.745 97.901 1.00 2.00 0 ATOM 1419 CB ILE 146 0.168 68.278 98.048 1.002.00 0 ATOM 1420 CG2 ILE 146 −0.607 68.690 99.278 1.00 2.00 0 ATOM 1421CG1 ILE 146 1.591 69.798 98.201 1.00 2.00 0 ATOM 1422 CD1 ILE 146 1.67970.270 99.360 1.00 2.00 0 ATOM 1423 C ILE 146 −1.181 66.143 97.851 1.002.00 0 ATOM 1424 O ILE 146 −1.805 65.927 98.881 1.00 2.00 0 ATOM 1425 NLYS 147 −1.680 65.839 96.668 1.00 2.00 0 ATOM 1427 CA LYS 147 −3.00065.241 96.594 1.00 2.00 0 ATOM 1428 CB LYS 147 −3.412 65.007 95.131 1.0015.32 0 ATOM 1429 CG LYS 147 −4.880 64.643 94.902 1.00 21.64 0 ATOM 1430CD LYS 147 −5.024 63.248 94.280 1.00 29.48 0 ATOM 1431 CE LYS 147 −4.70462.128 95.300 1.00 25.79 0 ATOM 1432 NZ LYS 147 −4.388 60.777 94.7161.00 22.31 0 ATOM 1436 C LYS 147 −2.864 63.928 97.345 1.00 2.00 0 ATOM1437 O LYS 147 −3.652 63.630 96.233 1.00 8.30 0 ATOM 1438 N LEU 148−1.815 63.180 97.023 1.00 9.36 0 ATOM 1440 CA LEU 148 −1.582 61.89297.651 1.00 9.36 0 ATOM 1441 CB LEU 148 −0.360 61.202 97.037 1.00 2.00 0ATOM 1442 CG LEU 148 −0.207 59.721 97.415 1.00 2.00 0 ATOM 1443 CD1 LEU148 −1.398 58.910 96.924 1.00 2.00 0 ATOM 1444 CD2 LEU 148 1.078 59.16896.835 1.00 2.00 0 ATOM 1445 C LEU 148 −1.423 62.040 99.157 1.00 9.36 0ATOM 1446 O LEU 148 −2.097 61.351 99.899 1.00 2.00 0 ATOM 1447 N TRP 149−0.555 62.943 99.611 1.00 2.00 0 ATOM 1449 CA TRP 149 −0.360 63.166101.042 1.00 2.00 0 ATOM 1450 CB TRP 149 0.559 64.359 101.276 1.00 9.480 ATOM 1451 CG TRP 149 0.690 64.767 102.748 1.00 13.35 0 ATOM 1452 CD2TRP 149 0.095 65.915 103.387 1.00 9.36 0 ATOM 1453 CE2 TRP 149 0.51265.907 104.725 1.00 12.67 0 ATOM 1454 CE3 TRP 149 −0.751 66.946 102.9511.00 9.36 0 ATOM 1455 CD1 TRP 149 1.415 64.135 103.713 1.00 12.84 0 ATOM1456 NE1 TRP 149 1.315 64.811 104.895 1.00 14.40 0 ATOM 1458 CZ2 TRP 1490.119 66.880 105.632 1.00 10.82 0 ATOM 1459 CZ3 TRP 149 −1.138 67.912103.858 1.00 10.25 0 ATOM 1460 CH2 TRP 149 −0.702 67.870 105.182 1.0021.34 0 ATOM 1461 C TRP 149 −1.712 63.427 101.707 1.00 2.00 0 ATOM 1462O TRP 149 −2.095 62.726 102.647 1.00 9.36 0 ATOM 1463 N LYS 150 −2.42964.434 101.216 1.00 2.00 0 ATOM 1465 CA LYS 150 −3.756 64.777 101.7251.00 2.00 0 ATOM 1466 CB LYS 150 −4.392 65.810 100.819 1.00 25.82 0 ATOM1467 CG LYS 150 −3.695 67.135 100.844 1.00 27.78 0 ATOM 1468 CD LYS 150−4.145 67.981 99.676 1.00 24.78 0 ATOM 1469 CE LYS 150 −4.236 69.442100.053 1.00 28.33 0 ATOM 1470 NZ LYS 150 −5.243 69.673 101.132 1.0035.49 0 ATOM 1474 C LYS 150 −4.661 63.536 101.786 1.00 2.00 0 ATOM 1475O LYS 150 −5.468 63.397 102.701 1.00 28.71 0 ATOM 1476 N THR 151 −4.52562.638 100.809 1.00 2.00 0 ATOM 1478 CA THR 151 −5.315 61.403 100.7581.00 2.00 0 ATOM 1479 CB THR 151 −5.111 60.670 99.408 1.00 18.71 0 ATOM1480 OG1 THR 151 −5.491 61.532 98.332 1.00 21.97 0 ATOM 1482 CG2 THR 151−5.964 59.434 99.332 1.00 19.85 0 ATOM 1483 C THR 151 −4.920 60.487101.925 1.00 2.00 0 ATOM 1484 O THR 151 −5.760 59.773 102.489 1.00 14.710 ATOM 1485 N PHE 152 −3.645 60.529 102.297 1.00 2.00 0 ATOM 1487 CA PHE152 −3.161 59.729 103.398 1.00 2.00 0 ATOM 1488 CB PHE 152 −1.638 59.759103.502 1.00 13.78 0 ATOM 1489 CG PHE 152 −0.956 58.5B6 102.830 1.0013.78 0 ATOM 1490 CD1 PHE 152 0.034 58.792 101.864 1.00 13.78 0 ATOM1491 CD2 PHE 152 −1.289 57.277 103.178 1.00 13.78 0 ATOM 1492 CE1 PHE152 0.679 57.717 101.260 1.00 13.78 0 ATOM 1493 CE2 PHE 152 −0.64856.194 102.578 1.00 13.78 0 ATOM 1494 CZ PHE 152 0.340 56.417 101.6171.00 13.78 0 ATOM 1495 C PHE 152 −3.767 60.246 104.673 1.00 2.00 0 ATOM1496 O PHE 152 −4.380 59.470 105.397 1.00 13.78 0 ATOM 1497 N THR 153−3.657 61.547 104.944 1.00 2.00 0 ATOM 1499 CA THR 153 −4.217 62.064106.192 1.00 2.00 0 ATOM 1500 CB THR 153 −4.166 63.604 106.318 1.0019.49 0 ATOM 1501 OG1 THR 153 −4.912 64.207 105.265 1.00 24.34 0 ATOM1503 CG2 THR 153 −2.739 64.095 106.293 1.00 23.93 0 ATOM 1504 C THR 153−5.653 61.007 106.348 1.00 2.00 0 ATOM 1505 O THR 153 −6.054 61.158107.414 1.00 29.22 0 ATOM 1506 N ASP 154 −6.415 61.667 105.276 1.00 2.000 ATOM 1508 CA ASP 154 −7.801 61.250 105.332 1.00 2.00 0 ATOM 1509 CBASP 154 −8.442 61.393 103.945 1.00 21.48 0 ATOM 1510 CG ASP 154 −9.96561.345 103.987 1.00 21.21 0 ATOM 1511 OD1 ASP 154 −10.576 61.750 102.9711.00 27.37 0 ATOM 1512 OD2 ASP 154 −10.548 60.912 105.018 1.00 20.95 0ATOM 1513 C ASP 154 −7.889 59.806 105.801 1.00 2.00 0 ATOM 1514 O ASP154 −8.783 59.444 106.569 1.00 11.19 0 ATOM 1515 N CYS 155 −6.968 58.981105.319 1.00 2.00 0 ATOM 1517 CA CYS 155 −6.951 57.571 105.691 1.00 2.000 ATOM 1518 CB CYS 155 −5.904 56.822 104.865 1.00 2.00 0 ATOM 1519 SGCYS 155 −5.770 55.072 105.254 1.00 2.00 0 ATOM 1520 C CYS 155 −6.62757.479 107.178 1.00 2.00 0 ATOM 1521 O CYS 155 −7.267 56.741 107.9311.00 2.00 0 ATOM 1522 N PHE 156 −5.641 58.273 107.593 1.00 8.58 0 ATOM1524 CA PHE 156 −5.172 58.366 108.954 1.00 8.58 0 ATOM 1525 CB PHE 156−4.056 59.409 109.018 1.00 2.00 0 ATOM 1526 CG PHE 156 −2.766 58.945108.437 1.00 2.00 0 ATOM 1527 CD1 PHE 156 −2.537 57.585 108.206 1.002.00 0 ATOM 1528 CD2 PHE 156 −1.756 59.847 108.160 1.00 2.00 0 ATOM 1529CE1 PHE 156 −1.309 57.130 107.710 1.00 2.00 0 ATOM 1530 CE2 PHE 156−0.518 59.400 107.662 1.00 2.00 0 ATOM 1531 CZ PHE 156 −0.295 58.039107.439 1.00 2.00 0 ATOM 1532 C PHE 156 −6.287 58.743 109.931 1.00 8.580 ATOM 1533 O PHE 156 −6.463 58.119 110.974 1.00 2.00 0 ATOM 1534 N ASN157 −7.055 59.758 109.572 1.00 2.00 0 ATOM 1536 CA ASN 157 −8.144 60.233110.405 1.00 2.00 0 ATOM 1537 CB ASN 157 −8.811 61.475 109.779 1.00 2.000 ATOM 1538 CG ASN 157 −7.861 62.661 109.577 1.00 2.00 0 ATOM 1539 OD1ASN 157 −8.226 63.629 108.922 1.00 2.00 0 ATOM 1540 ND2 ASN 157 −6.66962.601 110.143 1.00 2.00 0 ATOM 1543 C ASN 157 −9.230 59.179 110.6311.00 2.00 0 ATOM 1544 O ASN 157 −10.242 59.489 111.240 1.00 2.00 0 ATOM1545 N CYS 158 −9.064 57.962 110.120 1.00 2.00 0 ATOM 1547 CA CYS 158−10.074 56.914 110.304 1.00 2.00 0 ATOM 1548 CB CYS 158 −10.751 56.567108.970 1.00 2.00 0 ATOM 1549 SG CYS 158 −11.898 57.864 108.331 1.002.00 0 ATOM 1550 C CYS 158 −9.482 55.665 110.924 1.00 2.00 0 ATOM 1551 OCYS 158 −10.148 54.648 111.049 1.00 2.00 0 ATOM 1552 N LEU 159 −8.21655.772 111.304 1.00 2.00 0 ATOM 1554 CA LEU 159 −7.433 54.714 111.9451.00 2.00 0 ATOM 1555 CB LEU 159 −5.985 55.219 112.037 1.00 2.00 0 ATOM1556 CG LEU 159 −4.747 54.363 111.781 1.00 2.00 0 ATOM 1557 CD1 LEU 159−4.993 53.449 110.617 1.00 2.00 0 ATOM 1558 CD2 LEU 159 −3.536 55.269111.503 1.00 2.00 0 ATOM 1559 C LEU 159 −8.010 54.454 113.361 1.00 2.000 ATOM 1560 O LEU 159 −8.548 55.367 113.980 1.00 2.00 0 ATOM 1561 N PRO160 −7.951 53.209 113.871 1.00 2.00 0 ATOM 1562 CD PRO 160 −7.504 51.978113.208 1.00 2.00 0 ATOM 1563 CA PRO 160 −8.473 52.896 115.209 1.00 2.000 ATOM 1564 CB PRO 160 −8.398 51.375 115.269 1.00 2.00 0 ATOM 1565 CGPRO 160 −8.416 50.956 113.838 1.00 2.00 0 ATOM 1566 C PRO 160 −7.53553.516 116.228 1.00 2.00 0 ATOM 1567 O PRO 160 −6.329 53.463 116.0311.00 2.00 0 ATOM 1568 N ILE 161 −8.062 54.073 117.315 1.00 10.42 0 ATOM1570 CA ILE 161 −7.206 54.717 118.312 1.00 10.42 0 ATOM 1571 CB ILE 161−7.862 55.995 118.918 1.00 2.00 0 ATOM 1572 CG2 ILE 161 −8.493 56.837117.823 1.00 2.00 0 ATOM 1573 CG1 ILE 161 −8.899 55.612 119.987 1.002.00 0 ATOM 1574 CD1 ILE 161 −9.428 56.766 120.775 1.00 2.00 0 ATOM 1575C ILE 161 −6.741 53.859 119.492 1.00 10.42 0 ATOM 1576 O ILE 161 −5.80854.254 120.193 1.00 2.00 0 ATOM 1577 N ALA 162 −7.387 52.712 119.7201.00 16.67 0 ATOM 1579 CA ALA 162 −7.036 51.832 120.837 1.00 16.67 0ATOM 1580 CB ALA 162 −7.580 52.409 122.120 1.00 2.00 0 ATOM 1581 C ALA162 −7.567 50.414 120.637 1.00 16.67 0 ATOM 1582 O ALA 162 −8.494 50.203119.861 1.00 2.00 0 ATOM 1583 N ALA 163 −6.984 49.455 121.356 1.00 2.000 ATOM 1585 CA ALA 163 −7.379 48.052 121.267 1.00 2.00 0 ATOM 1586 CBALA 163 −6.559 47.374 120.197 1.00 14.80 0 ATOM 1587 C ALA 163 −7.23247.283 122.604 1.00 2.00 0 ATOM 1588 O ALA 163 −6.373 47.620 123.4251.00 8.24 0 ATOM 1589 N ILE 164 −8.069 46.263 122.813 1.00 8.09 0 ATOM1591 CA ILE 164 −8.036 45.424 124.018 1.00 8.09 0 ATOM 1592 CB ILE 164−9.323 45.590 124.860 1.00 10.56 0 ATOM 1593 CG2 ILE 164 −9.200 44.830126.150 1.00 10.56 0 ATOM 1594 CG1 ILE 164 −9.566 47.055 125.187 1.0010.56 0 ATOM 1595 CD1 ILE 164 −10.886 47.285 125.837 1.00 10.56 0 ATOM1596 C ILE 164 −7.910 43.938 123.623 1.00 8.09 0 ATOM 1597 O ILE 164−8.866 43.328 123.127 1.00 10.56 0 ATOM 1598 N VAL 165 −6.739 43.353123.856 1.00 20.53 0 ATOM 1600 CA VAL 165 −6.510 41.958 123.503 1.0021.83 0 ATOM 1601 CB VAL 165 −5.041 41.664 123.243 1.00 2.00 0 ATOM 1602CG1 VAL 165 −4.905 40.241 122.717 1.00 2.00 0 ATOM 1603 CG2 VAL 165−4.460 42.686 122.273 1.00 2.00 0 ATOM 1604 C VAL 165 −6.973 40.987124.570 1.00 22.04 0 ATOM 1605 O VAL 165 −6.546 41.064 125.728 1.00 2.000 ATOM 1606 N ASP 166 −7.841 40.066 124.163 1.00 10.50 0 ATOM 1608 CAASP 166 −8.401 39.057 125.046 1.00 16.73 0 ATOM 1609 CB ASP 166 −7.34837.980 125.337 1.00 24.26 0 ATOM 1610 CG ASP 166 −7.245 36.914 124.2121.00 24.02 0 ATOM 1611 OD1 ASP 166 −8.258 36.207 123.944 1.00 22.39 0ATOM 1612 OD2 ASP 166 −6.145 36.776 123.611 1.00 26.91 0 ATOM 1613 C ASP166 −8.963 39.674 126.326 1.00 15.15 0 ATOM 1614 O ASP 166 −9.139 39.006127.336 1.00 12.89 0 ATOM 1615 N GLU 167 −9.262 40.967 126.231 1.00 2.000 ATOM 1617 CA GLU 167 −9.833 41.811 127.289 1.00 2.00 0 ATOM 1618 CBGLU 167 −11.280 41.394 127.555 1.00 83.32 0 ATOM 1619 CG GLU 167 −12.12941.397 126.273 1.00 2.00. 0 ATOM 1620 CD GLU 167 −11.819 42.603 125.3051.00 2.00 0 ATOM 1621 OE1 GLU 167 −11.133 42.398 124.242 1.00 2.00 0ATOM 1622 OE2 GLU 167 −12.268 43.750 125.622 1.00 2.00 0 ATOM 1623 C GLU167 −9.056 41.981 128.585 1.00 2.00 0 ATOM 1624 O GLU 167 −9.634 42.138129.657 1.00 74.79 0 ATOM 1625 N LYS 168 −7.733 41.984 128.460 1.0022.77 0 ATOM 1627 CA LYS 168 −6.829 42.154 129.589 1.00 14.49 0 ATOM1628 CB LYS 168 −6.098 40.844 129.910 1.00 21.81 0 ATOM 1629 CG LYS 168−6.956 39.798 130.600 1.00 17.49 0 ATOM 1630 CD LYS 168 −7.677 40.404131.802 1.00 18.27 0 ATOM 1631 CE LYS 168 −8.654 39.411 132.466 1.0026.28 0 ATOM 1632 NZ LYS 168 −9.658 40.080 133.373 1.00 29.04 0 ATOM1636 C LYS 166 −5.811 43.225 129.237 1.00 11.04 0 ATOM 1637 O LYS 168−5.665 44.214 129.947 1.00 17.06 0 ATOM 1638 N ILE 169 −5.109 43.015128.131 1.00 2.00 0 ATOM 1640 CA ILE 169 −4.093 43.950 127.663 1.00 2.000 ATOM 1641 CB ILE 169 −3.151 43.267 126.648 1.00 2.00 0 ATOM 1642 CG2ILE 169 −1.989 44.184 126.302 1.00 2.00 0 ATOM 1643 CG1 ILE 169 −2.63241.952 127.215 1.00 2.00 0 ATOM 1644 CD1 ILE 169 −1.691 41.218 126.2971.00 2.00 0 ATOM 1645 C ILE 169 −4.731 45.144 126.964 1.00 2.00 0 ATOM1646 O ILE 169 −5.393 44.973 125.938 1.00 2.00 0 ATOM 1647 N PHE 170−4.572 46.337 127.524 1.00 17.81 0 ATOM 1649 CA PHE 170 −5.110 47.525126.877 1.00 19.78 0 ATOM 1650 CB PHE 170 −5.486 48.610 127.885 1.002.00 0 ATOM 1651 CG PHE 170 −5.895 49.920 127.252 1.00 2.00 0 ATOM 1652CD1 PHE 170 −7.095 50.028 126.538 1.00 2.00 0 ATOM 1653 CD2 PHE 170−5.092 51.056 127.390 1.00 2.00 0 ATOM 1654 CE1 PHE 170 −7.493 51.248125.976 1.00 2.00 0 ATOM 1655 CE2 PHE 170 −5.473 52.282 126.836 1.002.00 0 ATOM 1656 CZ PHE 170 −6.680 52.377 126.127 1.00 2.00 0 ATOM 1657C PHE 170 −3.943 47.992 126.038 1.00 23.45 0 ATOM 1658 O PHE 170 −2.80947.983 126.518 1.00 2.00 0 ATOM 1659 N CYS 171 −4.208 48.375 124.7901.00 2.00 0 ATOM 1661 CA CYS 171 −3.158 48.836 123.868 1.00 2.00 0 ATOM1662 CB CYS 171 −2.952 47.643 122.766 1.00 26.14 0 ATOM 1663 SG CYS 171−2.524 46.203 123.305 1.00 16.13 0 ATOM 1664 C CYS 171 −3.499 50.189123.310 1.00 2.00 0 ATOM 1665 O CYS 171 −4.652 50.453 122.943 1.00 26.140 ATOM 1666 N CYS 172 −2.495 51.058 123.265 1.00 2.00 0 ATOM 1668 CA CYS172 −2.635 52.378 122.668 1.00 2.00 0 ATOM 1669 CB CYS 172 −3.403 53.356123.597 1.00 14.54 0 ATOM 1670 SG CYS 172 −2.593 53.995 125.094 1.0015.67 0 ATOM 1671 C CYS 172 −1.231 52.890 122.287 1.00 2.00 0 ATOM 1672O CYS 172 −0.227 52.277 122.657 1.00 7.48 0 ATOM 1673 N HIS 173 −1.15453.951 121.488 1.00 2.00 0 ATOM 1675 CA HIS 173 0.146 54.461 121.1121.00 2.00 0 ATOM 1676 C HIS 173 0.815 55.130 122.297 1.00 2.00 0 ATOM1677 O HIS 173 1.893 54.707 122.722 1.00 2.00 0 ATOM 1678 CB HIS 1730.056 55.457 119.944 1.00 2.00 0 ATOM 1679 CG HIS 173 1.377 56.074119.588 1.00 2.00 0 ATOM 1680 ND1 HIS 173 2.487 55.347 119.234 1.00 2.000 ATOM 1682 CD2 HIS 173 1.774 57.372 119.565 1.00 2.00 0 ATOM 1683 NE2HIS 173 3.123 57.441 119.241 1.00 2.00 0 ATOM 1684 CE1 HIS 173 3.49256.189 119.045 1.00 2.00 0 ATOM 1685 N GLY 174 0.168 56.172 122.817 1.002.00 0 ATOM 1697 CA GLY 174 0.711 56.933 123.935 1.00 2.00 0 ATOM 1688 CGLY 174 0.568 56.450 125.386 1.00 2.00 0 ATOM 1689 O GLY 174 1.55656.111 126.041 1.00 2.00 0 ATOM 1690 N GLY 175 −0.648 56.453 125.9131.00 16.98 0 ATOM 1692 CA GLY 175 −0.830 56.028 127.283 1.00 16.98 0ATOM 1693 C GLY 175 −2.227 56.343 127.763 1.00 16.98 0 ATOM 1694 O GLY175 −3.190 56.231 126.999 1.00 66.77 0 ATOM 1695 N LEU 176 −2.341 56.764129.021 1.00 11.28 0 ATOM 1697 CA LEU 176 −3.637 57.063 129.604 1.0011.28 0 ATOM 1698 CB LEU 176 −3.740 56.427 130.976 1.00 2.00 0 ATOM 1699CG LEU 176 −3.443 54.934 130.966 1.00 2.00 0 ATOM 1700 CD1 LEU 176−3.469 54.407 132.374 1.00 2.00 0 ATOM 1701 CD2 LEU 176 −4.463 54.218130.121 1.00 2.00 0 ATOM 1702 C LEU 176 −3.876 58.545 129.692 1.00 11.280 ATOM 1703 O LEU 176 −2.943 59.329 129.645 1.00 2.00 0 ATOM 1704 N SER177 −5.138 58.913 129.850 1.00 6.18 0 ATOM 1706 CA SER 177 −5.567 60.304129.918 1.00 6.86 0 ATOM 1707 CB SER 177 −6.476 60.587 128.718 1.0012.19 0 ATOM 1708 OG SER 177 −7.189 61.804 128.835 1.00 12.19 0 ATOM1710 C SER 177 −6.356 60.573 131.193 1.00 7.79 0 ATOM 1711 O SER 177−7.170 59.749 131.606 1.00 12.19 0 ATOM 1712 N PRO 178 −6.150 61.736131.824 1.00 2.00 0 ATOM 1713 CD PRO 178 −5.223 62.827 131.505 1.0029.89 0 ATOM 1714 CA PRO 178 −6.895 62.054 133.041 1.00 2.00 0 ATOM 1715CB PRO 178 −6.231 63.337 133.518 1.00 25.57 0 ATOM 1716 CG PRO 178−5.842 63.973 132.274 1.00 21.36 0 ATOM 1717 C PRO 178 −8.394 62.266132.757 1.00 2.00 0 ATOM 1718 O PRO 178 −9.140 62.740 133.617 1.00 24.040 ATOM 1719 N ASP 179 −8.821 61.933 131.543 1.00 38.94 0 ATOM 1721 CAASP 179 −10.206 62.078 131.129 1.00 37.05 0 ATOM 1722 CB ASP 179 −10.26462.870 129.837 1.00 33.34 0 ATOM 1723 CG ASP 179 −9.964 64.320 130.0481.00 30.55 0 ATOM 1724 OD1 ASP 179 −10.923 65.045 130.386 1.00 31.71 0ATOM 1725 OD2 ASP 179 −8.796 64.728 129.889 1.00 32.17 0 ATOM 1726 C ASP179 −10.871 60.735 130.923 1.00 40.05 0 ATOM 1727 O ASP 179 −12.09660.638 130.873 1.00 34.46 0 ATOM 1728 N LEU 180 −10.057 59.699 130.7911.00 15.43 0 ATOM 1730 CA LEU 180 −10.570 58.362 130.590 1.00 12.98 0ATOM 1731 CB LEU 180 −9.446 57.467 130.058 1.00 2.00 0 ATOM 1732 CG LEU180 −9.867 56.186 129.336 1.00 2.00 0 ATOM 1733 CD1 LEU 180 −10.70656.511 128.110 1.00 2.00 0 ATOM 1734 CD2 LEU 180 −8.633 55.413 128.9531.00 2.00 0 ATOM 1735 C LEU 180 −11.157 57.806 131.902 1.00 14.43 0 ATOM1736 O LEU 180 −10.470 57.709 132.931 1.00 2.00 0 ATOM 1737 N GLN 181−12.448 57.492 131.862 1.00 5.88 0 ATOM 1739 CA GLN 181 −13.184 56.917132.992 1.00 5.88 0 ATOM 1740 CB GLN 181 −14.379 57.787 133.375 1.0032.99 0 ATOM 1741 CG GLN 181 −14.002 59.152 133.900 1.00 36.07 0 ATOM1742 CD GLN 181 −15.101 60.172 133.676 1.00 38.35 0 ATOM 1743 OE1 GLN181 −16.285 59.886 133.877 1.00 41.53 0 ATOM 1744 NE2 GLN 181 −14.71761.366 133.242 1.00 38.39 0 ATOM 1747 C GLN 181 −13.672 55.563 132.5001.00 5.88 0 ATOM 1748 O GLN 181 −13.572 54.554 133.205 1.00 24.31 0 ATOM1749 N SER 182 −14.188 55.544 131.274 1.00 2.20 0 ATOM 1751 CA SER 182−14.651 54.310 130.664 1.00 9.71 0 ATOM 1752 CB SER 182 −16.174 54.255130.614 1.00 15.80 0 ATOM 1753 OG SER 182 −16.668 54.856 129.433 1.0017.25 0 ATOM 1755 C SER 182 −14.112 54.177 129.248 1.00 3.77 0 ATOM 1756O SER 182 −13.359 55.019 126.751 1.00 15.80 0 ATOM 1757 N MET 183−14.526 53.091 126.615 1.00 2.00 0 ATOM 1759 CA MET 163 −14.156 52.780127.251 1.00 2.00 0 ATOM 1760 CB MET 183 −14.100 51.272 127.064 1.002.00 0 ATOM 1761 CG MET 183 −13.171 50.616 128.048 1.00 2.00 0 ATOM 1762SD MET 183 −11.620 51.473 128.022 1.00 2.00 0 ATOM 1763 CE MET 183−10.520 50.135 127.900 1.00 2.00 0 ATOM 1764 C MET 183 −15.204 53.373126.326 1.00 2.00 0 ATOM 1765 O MET 183 −14.959 53.550 125.129 1.00 2.000 ATOM 1766 N GLU 184 −16.370 53.700 126.882 1.00 2.00 0 ATOM 1768 CAGLU 184 −17.432 54.280 126.082 1.00 2.00 0 ATOM 1769 CB GLU 184 −18.66854.531 126.910 1.00 6.25 0 ATOM 1770 CG GLU 184 −19.830 54.977 126.0731.00 8.82 0 ATOM 1771 CD GLU 184 −20.273 53.927 125.068 1.00 13.01 0ATOM 1772 OE1 GLU 184 −19.846 52.744 125.181 1.00 14.09 0 ATOM 1773 OE2GLU 184 −21.064 54.293 124.164 1.00 20.29 0 ATOM 1774 C GLU 184 −16.97855.586 125.481 1.00 2.00 0 ATOM 1775 O GLU 184 −17.399 55.946 124.3921.00 23.33 0 ATOM 1776 N GLN 185 −16.117 56.295 126.198 1.00 12.32 0ATOM 1778 CA GLN 185 −15.599 57.561 125.714 1.00 14.30 0 ATOM 1779 CBGLN 185 −14.697 58.190 126.758 1.00 43.49 0 ATOM 1780 CG GLN 185 −15.45458.629 127.990 1.00 47.40 0 ATOM 1781 CD GLN 185 −14.537 58.929 129.1391.00 49.32 0 ATOM 1782 OE1 GLN 185 −13.994 58.016 129.753 1.00 56.49 0ATOM 1783 NE2 GLN 185 −14.350 60.208 129.437 1.00 49.76 0 ATOM 1786 CGLN 185 −14.834 57.307 124.432 1.00 16.93 0 ATOM 1787 O GLN 185 −14.97358.053 123.461 1.00 43.79 0 ATOM 1788 N ILE 186 −14.044 56.238 124.4201.00 5.67 0 ATOM 1790 CA ILE 186 −13.280 55.880 123.235 1.00 5.67 0 ATOM1791 CB ILE 186 −12.436 54.613 123.470 1.00 16.18 0 ATOM 1792 CG2 ILE186 −11.675 54.243 122.208 1.00 11.85 0 ATOM 1793 CG1 ILE 186 −11.48954.825 124.642 1.00 13.18 0 ATOM 1794 CD1 ILE 186 −10.545 55.954 124.4591.00 17.92 0 ATOM 1795 C ILE 186 −14.294 55.566 122.143 1.00 5.67 0 ATOM1796 O ILE 186 −14.260 56.145 121.060 1.00 19.35 0 ATOM 1797 N ARG 187−15.207 54.655 122.471 1.00 17.59 0 ATOM 1799 CA ARG 187 −16.243 54.205121.561 1.00 16.33 0 ATOM 1800 CB ARG 187 −17.141 53.165 122.237 1.0020.41 0 ATOM 1801 CG ARG 187 −16.468 51.900 122.723 1.00 29.32 0 ATOM1802 CD ARG 187 −17.497 51.006 123.447 1.00 31.43 0 ATOM 1803 NE ARG 187−16.888 49.888 124.165 1.00 38.34 0 ATOM 1805 CZ ARG 187 −16.311 48.838123.580 1.00 34.30 0 ATOM 1806 NH1 ARG 187 −16.256 48.747 122.252 1.0041.37 0 ATOM 1809 NH2 ARG 187 −15.783 47.873 124.322 1.00 34.50 0 ATOM1812 C ARG 187 −17.148 55.302 121.011 1.00 15.89 0 ATOM 1813 O ARG 187−17.937 55.032 120.108 1.00 20.82 0 ATOM 1814 N ARG 188 −17.071 56.524121.529 1.00 2.00 0 ATOM 1816 CA ARG 188 −17.964 57.557 121.012 1.002.00 0 ATOM 1817 CE ARG 188 −18.878 58.105 122.106 1.00 31.38 0 ATOM1818 CG ARG 188 −18.184 56.827 123.228 1.00 29.44 0 ATOM 1819 CD ARG 188−19.202 59.586 124.034 1.00 31.32 0 ATOM 1820 NE ARG 188 −20.410 58.802124.237 1.00 33.18 0 ATOM 1822 CZ ARG 188 −21.637 59.285 124.112 1.0029.63 0 ATOM 1823 NH1 ARG 188 −21.827 60.561 123.777 1.00 36.26 0 ATOM1826 NH2 ARG 188 −23.671 58.484 124.332 1.00 32.06 0 ATOM 1829 C ARG 188−17.255 58.692 120.332 1.00 2.00 0 ATOM 1830 O ARG 188 −17.758 59.904120.250 1.00 31.89 0 ATOM 1831 N ILE 189 −16.070 58.369 119.846 1.0017.06 0 ATOM 1833 CA ILE 189 −15.263 59.333 119.136 1.00 16.98 0 ATOM1834 CB ILE 189 −13.759 59.043 119.345 1.00 16.67 0 ATOM 1835 CG2 ILE189 −12.924 59.973 118.493 1.00 17.43 0 ATOM 1836 CG1 ILE 189 −13.41159.205 120.826 1.00 17.65 0 ATOM 1837 CD1 ILE 189 −11.970 58.946 121.1731.00 16.74 0 ATOM 1838 C ILE 189 −15.620 59.244 117.656 1.00 19.51 0ATOM 1839 O ILE 189 −15.448 58.184 117.044 1.00 25.93 0 ATOM 1840 N MET190 −16.158 60.342 117.108 1.00 22.12 0 ATOM 1842 CA MET 190 −16.51460.420 115.685 1.00 24.92 0 ATOM 1843 CB MET 190 −17.051 61.812 115.3201.00 46.39 0 ATOM 1844 CG MET 190 −17.366 61.977 113.826 1.00 46.97 0ATOM 1845 SD MET 190 −17.820 63.658 113.312 1.00 58.03 0 ATOM 1846 CEMET 190 −16.271 64.266 112.598 1.00 52.38 0 ATOM 1847 C MET 190 −15.17360.194 115.015 1.00 25.36 0 ATOM 1848 O MET 190 −14.287 61.041 115.1061.00 32.57 0 ATOM 1849 N ARG 191 −15.023 59.057 114.346 1.00 2.00 0 ATOM1851 CA ARG 191 −13.732 58.738 113.772 1.00 2.00 0 ATOM 1852 CB ARG 191−13.731 57.394 113.089 1.00 2.00 0 ATOM 1853 CG ARG 191 −12.300 56.900112.821 1.00 11.31 0 ATOM 1854 CD ARG 191 −11.978 55.799 113.752 1.0011.31 0 ATOM 1855 NE ARG 191 −13.139 54.935 113.762 1.00 2.00 0 ATOM1857 CZ ARG 191 −13.124 53.663 113.416 1.00 2.00 0 ATOM 1858 NH1 ARG 191−11.985 53.085 113.047 1.00 2.00 0 ATOM 1861 NH2 ARG 191 −14.263 52.984113.430 1.00 2.00 0 ATOM 1864 C ARG 191 −13.084 59.769 112.857 1.00 2.000 ATOM 1865 O ARG 191 −11.995 60.267 113.202 1.00 6.10 0 ATOM 1866 N PRO192 −13.685 60.065 111.670 1.00 37.06 0 ATOM 1867 CD PRO 192 −14.91259.530 111.045 1.00 2.00 0 ATOM 1868 CA PRO 192 −13.048 61.068 110.7991.00 36.28 0 ATOM 1869 CB PRO 192 −14.114 61.335 109.749 1.00 2.00 0ATOM 1870 CG PRO 192 −14.743 59.973 109.602 1.00 2.00 0 ATOM 1871 C PRO192 −12.787 62.261 111.693 1.00 37.24 0 ATOM 1872 O PRO 192 −13.69763.033 112.004 1.00 2.00 0 ATOM 1873 N THR 193 −11.552 62.348 112.1681.00 2.00 0 ATOM 1875 CA THR 193 −11.182 63.396 113.089 1.00 2.00 0 ATOM1876 CB THR 193 −11.627 63.063 114.540 1.00 22.05 0 ATOM 1877 OG1 THR193 −11.349 64.183 115.392 1.00 31.69 0 ATOM 1879 CG2 THR 193 −10.86261.859 115.089 1.00 21.46 0 ATOM 1880 C THR 193 −9.700 63.661 113.1521.00 2.00 0 ATOM 1881 O THR 193 −8.865 62.795 112.860 1.00 17.62 0 ATOM1882 N ASP 194 −9.394 64.879 113.572 1.00 2.00 0 ATOM 1884 CA ASP 194−8.031 65.311 113.744 1.00 2.00 0 ATOM 1885 CB ASP 194 −7.927 66.805113.431 1.00 49.25 0 ATOM 1886 CG ASP 194 −6.561 67.197 112.912 1.0049.25 0 ATOM 1887 OD1 ASP 194 −6.132 68.334 113.194 1.00 49.25 0 ATOM1888 OD2 ASP 194 −5.920 66.371 112.217 1.00 49.25 0 ATOM 1889 C ASP 194−7.686 65.041 115.216 1.00 2.00 0 ATOM 1890 O ASP 194 −8.571 64.978116.077 1.00 54.62 0 ATOM 1891 N VAL 195 −6.408 64.836 115.488 1.0017.01 0 ATOM 1893 CA VAL 195 −5.948 64.611 116.839 1.00 12.88 0 ATOM1894 CB VAL 195 −4.509 64.017 116.818 1.00 9.98 0 ATOM 1895 CG1 VAL 195−3.934 63.875 118.239 1.00 9.98 0 ATOM 1896 CG2 VAL 195 −4.532 62.675116.124 1.00 9.98 0 ATOM 1897 C VAL 195 −5.971 65.994 117.511 1.00 12.880 ATOM 1898 O VAL 195 −5.343 66.934 117.038 1.00 9.98 0 ATOM 1899 N PRO196 −6.771 66.152 118.567 1.00 16.08 0 ATOM 1900 CD PRO 196 −7.80565.214 119.033 1.00 26.02 0 ATOM 1901 CA PRO 196 −6.862 67.429 119.2841.00 21.91 0 ATOM 1902 CB PRO 196 −8.026 67.195 120.240 1.00 25.86 0ATOM 1903 CG PRO 196 −8.874 66.153 119.496 1.00 25.31 0 ATOM 1904 C PRO196 −5.560 67.735 120.027 1.00 24.64 0 ATOM 1905 O PRO 196 −4.809 66.818120.349 1.00 23.54 0 ATOM 1906 N ASP 197 −5.291 69.010 120.302 1.0039.93 0 ATOM 1908 CA ASP 197 −4.058 69.377 121.005 1.00 38.76 0 ATOM1909 CB ASP 197 −3.605 70.789 120.612 1.00 78.93 0 ATOM 1910 CG ASP 197−4.726 71.800 120.658 1.00 84.50 0 ATOM 1911 OD1 ASP 197 −5.071 72.263121.768 1.00 83.41 0 ATOM 1912 OD2 ASP 197 −5.258 72.134 119.578 1.0090.03 0 ATOM 1913 C ASP 197 −4.162 69.256 122.520 1.00 41.64 0 ATOM 1914O ASP 197 −3.227 69.577 123.255 1.00 85.35 0 ATOM 1915 N GLN 198 −5.31668.789 122.973 1.00 2.00 0 ATOM 1917 CA GLN 198 −5.581 66.588 124.3851.00 2.00 0 ATOM 1918 CB GLN 198 −5.686 69.937 125.120 1.00 27.32 0 ATOM1919 CG GLN 198 −6.621 70.967 124.503 1.00 32.39 0 ATOM 1920 CD GLN 198−7.981 71.041 125.192 1.00 29.94 0 ATOM 1921 OE1 GLN 198 −8.108 70.793126.397 1.00 32.81 0 ATOM 1922 NE2 GLN 198 −9.007 71.391 124.424 1.0029.42 0 ATOM 1925 C GLN 198 −6.863 67.793 124.492 1.00 2.00 0 ATOM 1926O GLN 198 −7.748 67.909 123.640 1.00 26.90 0 ATOM 1927 N GLY 199 −6.94366.949 125.509 1.00 15.20 0 ATOM 1929 CA GLY 199 −8.137 66.148 125.6921.00 15.56 0 ATOM 1930 C GLY 199 −7.820 64.670 125.631 1.00 13.32 0 ATOM1931 O GLY 199 −6.655 64.283 125.502 1.00 2.00 0 ATOM 1932 N LEU 200−8.857 63.842 125.684 1.00 2.00 0 ATOM 1934 CA LEU 200 −8.690 62.402125.666 1.00 2.00 0 ATOM 1935 CB LEU 200 −10.049 61.701 125.667 1.002.00 0 ATOM 1936 CG LEU 200 −9.945 60.224 126.081 1.00 2.00 0 ATOM 1937CD1 LEU 200 −11.126 59.800 126.947 1.00 2.00 0 ATOM 1938 CD2 LEU 200−9.842 59.380 124.847 1.00 2.00 0 ATOM 1939 C LEU 200 −7.850 61.875124.521 1.00 2.00 0 ATOM 1940 O LEU 200 −6.764 61.323 124.764 1.00 2.000 ATOM 1941 N LEU 201 −8.344 62.045 123.283 1.00 2.00 0 ATOM 1943 CA LEU201 −7.663 61.566 122.070 1.00 2.00 0 ATOM 1944 CB LEU 201 −8.378 62.041120.811 1.00 2.00 0 ATOM 1945 CG LEU 201 −8.369 61.100 119.589 1.00 2.000 ATOM 1946 CD1 LEU 201 −8.895 61.855 118.375 1.00 2.00 0 ATOM 1947 CD2LEU 201 −6.981 60.569 119.277 1.00 2.00 0 ATOM 1948 C LEU 201 −6.24662.078 122.059 1.00 2.00 0 ATOM 1949 O LEU 201 −5.319 61.347 121.7151.00 2.00 0 ATOM 1950 N CYS 202 −6.071 63.333 122.446 1.00 2.00 0 ATOM1952 CA CYS 202 −4.733 63.891 122.484 1.00 2.00 0 ATOM 1953 CB CYS 202−4.722 65.306 123.024 1.00 2.00 0 ATOM 1954 SG CYS 202 −3.012 65.850123.209 1.00 2.00 0 ATOM 1955 C CYS 202 −3.850 63.076 123.394 1.00 2.000 ATOM 1956 O CYS 202 −2.719 62.715 123.036 1.00 2.00 0 ATOM 1957 N ASP203 −4.385 62.798 124.584 1.00 15.07 0 ATOM 1959 CA ASP 203 −3.67362.057 125.611 1.00 11.86 0 ATOM 1960 CB ASP 203 −4.429 62.142 126.9311.00 13.71 0 ATOM 1961 CG ASP 203 −4.449 63.538 127.497 1.00 15.77 0ATOM 1962 OD1 ASP 203 −3.529 64.315 127.162 1.00 11.73 0 ATOM 1963 OD2ASP 203 −5.380 63.861 128.271 1.00 16.16 0 ATOM 1964 C ASP 203 −3.36960.615 125.262 1.00 13.81 0 ATOM 1965 O ASP 203 −2.254 60.155 125.4921.00 23.94 0 ATOM 1966 N LEU 204 −4.330 59.902 124.695 1.00 10.49 0 ATOM1968 CA LEU 204 −4.097 58.509 124.332 1.00 10.49 0 ATOM 1969 CB LEU 204−5.351 57.912 123.717 1.00 10.22 0 ATOM 1970 CG LEU 204 −6.478 57.606124.696 1.00 10.22 0 ATOM 1971 CD1 LEU 204 −7.667 57.281 123.866 1.0010.22 0 ATOM 1972 CD2 LEU 204 −6.143 56.441 125.620 1.00 10.22 0 ATOM1973 C LEU 204 −2.930 58.347 123.361 1.00 10.49 0 ATOM 1974 O LEU 204−2.204 57.352 123.392 1.00 10.22 0 ATOM 1975 N LEU 205 −2.748 59.336122.501 1.00 2.00 0 ATOM 1977 CA LEU 205 −1.679 59.301 121.525 1.00 2.000 ATOM 1978 CB LEU 205 −2.161 59.933 120.219 1.00 2.00 0 ATOM 1979 CGLEU 205 −3.459 59.463 119.551 1.00 2.00 0 ATOM 1980 CD1 LEU 205 −3.59560.238 118.259 1.00 2.00 0 ATOM 1981 CD2 LEU 205 −3.459 57.954 119.2691.00 2.00 0 ATOM 1982 C LEU 205 −0.409 60.018 121.967 1.00 2.00 0 ATOM1983 O LEU 205 0.620 59.890 121.314 1.00 2.00 0 ATOM 1984 N TRP 206−0.472 60.756 123.072 1.00 10.02 0 ATOM 1986 CA TRP 206 0.678 61.526123.545 1.00 10.02 0 ATOM 1987 CB TRP 206 0.362 62.994 123.408 1.00 2.000 ATOM 1988 CG TRP 206 0.413 63.485 122.052 1.00 2.00 0 ATOM 1989 CD2TRP 206 1.532 64.103 121.420 1.00 2.00 0 ATOM 1990 CE2 TRP 206 1.10564.535 120.148 1.00 2.00 0 ATOM 1991 CE3 TRP 206 2.854 64.355 121.8131.00 2.00 0 ATOM 1992 CD1 TRP 206 −0.616 63.541 121.162 1.00 2.00 0 ATOM1993 NE1 TRP 206 −0.212 64.178 120.017 1.00 2.00 0 ATOM 1995 CZ2 TRP 2061.950 65.207 119.263 1.00 2.00 0 ATOM 1996 CZ3 TRP 206 3.697 65.024120.932 1.00 2.00 0 ATOM 1997 CH2 TRP 206 3.238 65.443 119.669 1.00 2.000 ATOM 1998 C TRP 206 1.246 61.330 124.955 1.00 10.02 0 ATOM 1999 0 TRP206 2.419 61.639 125.194 1.00 2.00 0 ATOM 2000 N SER 207 0.418 60.967125.886 1.00 2.00 0 ATOM 2002 CA SER 207 0.830 60.690 127.267 1.00 2.000 ATOM 2003 CB SER 207 −0.363 60.273 128.121 1.00 29.76 0 ATOM 2004 OGSER 207 −0.482 61.120 129.256 1.00 35.73 0 ATOM 2006 C SER 207 1.95159.699 127.446 1.00 2.00 0 ATOM 2007 O SER 207 2.075 58.743 126.681 1.0031.05 0 ATOM 2008 N ASP 208 2.777 59.943 128.462 1.00 2.00 0 ATOM 2010CA ASP 208 3.899 59.056 128.764 1.00 2.00 0 ATOM 2011 CB ASP 208 5.25759.714 128.505 1.00 42.39 0 ATOM 2012 CG ASP 208 5.296 60.532 127.2561.00 46.59 0 ATOM 2013 OD1 ASP 208 5.695 61.706 127.362 1.00 48.93 0ATOM 2014 OD2 ASP 208 4.959 60.008 126.180 1.00 44.70 0 ATOM 2015 C ASP208 3.903 58.705 130.231 1.00 2.00 0 ATOM 2016 O ASP 208 3.580 59.537131.072 1.00 45.54 0 ATOM 2017 N PRO 209 4.244 57.457 130.559 1.00 10.980 ATOM 2018 CD PRO 209 4.509 56.316 129.673 1.00 2.00 0 ATOM 2019 CA PRO209 4.299 57.040 131.957 1.00 10.98 0 ATOM 2020 CB PRO 209 4.407 55.522131.845 1.00 2.00 0 ATOM 2021 CG PRO 209 5.197 55.356 130.604 1.00 2.000 ATOM 2022 C PRO 209 5.584 57.675 132.507 1.00 10.98 0 ATOM 2023 O PRO209 6.448 58.078 131.712 1.00 2.00 0 ATOM 2024 N ASP 210 5.724 57.774133.833 1.00 2.00 0 ATOM 2026 CA ASP 210 6.931 58.360 134.413 1.00 2.000 ATOM 2027 CB ASP 210 6.755 59.863 134.614 1.00 31.00 0 ATOM 2028 CGASP 210 8.050 60.570 134.990 1.00 36.69 0 ATOM 2029 OD1 ASP 210 9.13259.956 134.874 1.00 33.32 0 ATOM 2030 OD2 ASP 210 7.989 61.756 135.3961.00 38.93 0 ATOM 2031 C ASP 210 7.304 57.734 135.729 1.00 2.00 0 ATOM2032 O ASP 210 6.448 57.320 136.496 1.00 28.60 0 ATOM 2033 N LYS 2118.608 57.666 135.966 1.00 4.23 0 ATOM 2035 CA LYS 211 9.170 57.136137.211 1.00 9.37 0 ATOM 2036 CB LYS 211 10.647 56.760 137.024 1.0017.66 0 ATOM 2037 CG LYS 211 10.941 55.441 136.345 1.00 29.60 0 ATOM2038 CD LYS 211 12.451 55.184 136.403 1.00 32.82 0 ATOM 2039 CE LYS 21112.852 53.831 135.809 1.00 42.70 0 ATOM 2040 NZ LYS 211 14.336 53.616135.852 1.00 41.45 0 ATOM 2044 C LYS 211 9.100 58.157 138.367 1.00 7.690 ATOM 2045 O LYS 211 8.746 57.803 139.481 1.00 18.85 0 ATOM 2046 N ASP212 9.456 59.414 138.093 1.00 2.00 0 ATOM 2048 CA ASP 212 9.468 60.483139.101 1.00 2.00 0 ATOM 2049 CB ASP 212 10.322 61.669 138.624 1.0039.78 0 ATOM 2050 CG ASP 212 11.483 61.254 137.754 1.00 46.31 0 ATOM2051 OD1 ASP 212 11.823 62.030 136.834 1.00 41.85 0 ATOM 2052 OD2 ASP212 12.051 60.166 137.987 1.00 46.28 0 ATOM 2053 C ASP 212 8.074 61.033139.412 1.00 2.00 0 ATOM 2054 O ASP 212 7.943 62.164 139.903 1.00 41.820 ATOM 2055 N VAL 213 7.040 60.250 139.124 1.00 2.00 0 ATOM 2057 CA VAL213 5.669 60.690 139.340 1.00 2.00 0 ATOM 2058 CB VAL 213 5.137 61.376138.050 1.00 2.00 0 ATOM 2059 CG1 VAL 213 3.652 61.247 137.922 1.00 2.000 ATOM 2060 CG2 VAL 213 5.505 62.826 138.076 1.00 2.00 0 ATOM 2061 C VAL213 4.767 59.526 139.763 1.00 2.00 0 ATOM 2062 O VAL 213 5.007 58.356139.390 1.00 2.00 0 ATOM 2063 N LEU 214 3.748 59.844 140.564 1.00 21.530 ATOM 2065 CA LEU 214 2.815 58.830 141.022 1.00 19.34 0 ATOM 2066 CBLEU 214 2.742 58.809 142.543 1.00 14.66 0 ATOM 2067 CG LEU 214 2.37157.438 143.097 1.00 15.86 0 ATOM 2068 CD1 LEU 214 3.507 56.477 142.8161.00 9.71 0 ATOM 2069 CD2 LEU 214 2.109 57.530 144.588 1.00 17.44 0 ATOM2070 C LEU 214 1.433 59.068 140.439 1.00 19.87 0 ATOM 2071 O LEU 2140.757 58.124 140.033 1.00 9.54 0 ATOM 2072 N GLY 215 1.003 60.322140.411 1.00 16.98 0 ATOM 2074 CA GLY 215 −0.299 60.640 139.852 1.0011.19 0 ATOM 2075 C GLY 215 −0.081 61.255 138.487 1.00 12.07 0 ATOM 2076O GLY 215 0.772 60.796 137.739 1.00 10.02 0 ATOM 2077 N TRP 216 −0.82462.303 138.167 1.00 2.00 0 ATOM 2079 CA TRP 216 −0.684 62.979 136.8901.00 2.00 0 ATOM 2080 CB TRP 216 −2.030 63.535 136.458 1.00 2.00 0 ATOM2081 CG TRP 216 −2.903 62.452 136.050 1.00 2.00 0 ATOM 2082 CD2 TRP 216−2.840 61.741 134.804 1.00 2.00 0 ATOM 2083 CE2 TRP 216 −3.806 60.713134.865 1.00 2.00 0 ATOM 2084 CE3 TRP 216 −2.050 61.865 133.648 1.002.00 0 ATOM 2085 CD1 TRP 216 −3.879 61.860 136.789 1.00 2.00 0 ATOM 2086NE1 TRP 216 −4.425 60.811 136.087 1.00 2.00 0 ATOM 2088 CZ2 TRP 216−4.005 59.812 133.809 1.00 2.00 0 ATOM 2089 CZ3 TRP 216 −2.251 60.970132.605 1.00 2.00 0 ATOM 2090 CH2 TRP 216 −3.217 59.961 132.695 1.002.00 0 ATOM 2091 C TRP 216 0.310 64.101 136.985 1.00 2.00 0 ATOM 2092 OTRP 216 0.261 64.883 137.926 1.00 2.00 0 ATOM 2093 N GLY 217 1.21364.182 136.021 1.00 12.63 0 ATOM 2095 CA GLY 217 2.199 65.245 136.0281.00 15.52 0 ATOM 2096 C GLY 217 2.312 65.928 134.675 1.00 10.15 0 ATOM2097 O GLY 217 1.627 65.559 133.720 1.00 26.84 0 ATOM 2098 N GLU 2183.189 66.923 134.594 1.00 23.86 0 ATOM 2100 CA GLU 218 3.434 67.686133.364 1.00 22.47 0 ATOM 2101 CB GLU 218 4.198 68.978 133.729 1.0065.82 0 ATOM 2102 CG GLU 218 5.082 69.595 132.641 1.00 67.96 0 ATOM 2103CD GLU 218 6.577 69.562 132.987 1.00 67.00 0 ATOM 2104 OE1 GLU 218 7.05670.519 133.631 1.00 68.41 0 ATOM 2105 OE2 GLU 218 7.276 68.589 132.6161.00 63.29 0 ATOM 2106 C GLU 218 4.232 66.841 132.371 1.00 25.48 0 ATOM2107 O GLU 218 4.808 65.820 132.750 1.00 67.07 0 ATOM 2108 N ASN 2194.253 67.245 131.103 1.00 21.19 0 ATOM 2110 CA ASN 219 5.039 66.510130.108 1.00 21.18 0 ATOM 2111 CB ASN 219 4.140 65.888 129.048 1.0010.60 0 ATOM 2112 CG ASN 219 4.832 64.791 128.282 1.00 14.78 0 ATOM 2113OD1 ASN 219 6.052 64.646 128.341 1.00 9.20 0 ATOM 2114 ND2 ASN 219 4.05764.003 127.560 1.00 8.28 0 ATOM 2117 C ASN 219 6.058 67.411 129.425 1.0021.70 0 ATOM 2118 O ASN 219 5.776 68.576 129.151 1.00 9.52 0 ATOM 2119 NASP 220 7.246 66.886 129.151 1.00 35.32 0 ATOM 2121 CA ASP 220 8.26667.690 128.485 1.00 34.19 0 ATOM 2122 CB ASP 220 9.585 66.900 128.3581.00 48.55 0 ATOM 2123 CG ASP 220 9.469 65.673 127.447 1.00 92.38 0 ATOM2124 OD1 ASP 220 9.083 64.584 127.938 1.00 48.43 0 ATOM 2125 OD2 ASP 2209.778 65.797 126.237 1.00 92.09 0 ATOM 2126 O ASP 220 7.763 68.148127.100 1.00 33.44 0 ATOM 2127 O ASP 220 8.045 69.262 126.661 1.00 47.840 ATOM 2128 N ARG 221 6.986 67.287 126.444 1.00 22.98 0 ATOM 2130 CA ARG221 6.427 67.552 125.127 1.00 22.11 0 ATOM 2131 CB ARG 221 5.578 66.382124.672 1.00 2.00 0 ATOM 2132 CG ARG 221 6.251 65.056 124.739 1.00 2.000 ATOM 2133 CD ARG 221 5.241 63.981 124.421 1.00 2.00 0 ATOM 2134 NE ARG221 5.828 62.658 124.590 1.00 2.00 0 ATOM 2136 CZ ARG 221 6.785 62.154123.811 1.00 2.00 0 ATOM 2137 NH1 ARG 221 7.255 62.866 122.790 1.00 2.000 ATOM 2140 NH2 ARG 221 7.285 60.947 124.064 1.00 2.00 0 ATOM 2143 C ARG221 5.546 68.776 125.109 1.00 18.96 0 ATOM 2144 O ARG 221 5.164 69.239124.052 1.00 2.00 0 ATOM 2145 N GLY 222 5.188 69.280 126.280 1.00 2.00 0ATOM 2147 CA GLY 222 4.328 70.447 126.343 1.00 2.00 0 ATOM 2148 C GLY222 2.939 70.103 125.844 1.00 2.00 0 ATOM 2149 O GLY 222 2.142 70.988125.509 1.00 29.15 0 ATOM 2150 N VAL 223 2.658 68.804 125.776 1.00 2.000 ATOM 2152 CA VAL 223 1.356 66.313 125.342 1.00 2.00 0 ATOM 2153 CB VAL223 1.364 67.860 123.839 1.00 2.00 0 ATOM 2154 CG1 VAL 223 −0.063 67.688123.280 1.00 2.00 0 ATOM 2155 CG2 VAL 223 2.258 68.762 123.004 1.00 2.000 ATOM 2156 C VAL 223 1.071 67.125 126.250 1.00 2.00 0 ATOM 2157 O VAL223 2.004 66.431 126.656 1.00 2.00 0 ATOM 2158 N SER 224 −0.205 66.897126.550 1.00 12.21 0 ATOM 2160 CA SER 224 −0.661 65.825 127.440 1.0015.55 0 ATOM 2161 CB SER 224 −0.609 64.453 126.759 1.00 6.64 0 ATOM 2162OG SER 224 0.713 64.003 126.546 1.00 10.80 0 ATOM 2164 C SER 224 0.07765.779 128.787 1.00 20.53 0 ATOM 2165 O SER 224 0.814 66.706 129.1531.00 8.42 0 ATOM 2166 N PHE 225 −0.124 64.700 129.533 1.00 6.68 0 ATOM2168 CA PHE 225 0.484 64.586 130.839 1.00 6.68 0 ATOM 2169 CB PHE 225−0.625 64.489 131.890 1.00 14.03 0 ATOM 2170 CG PHE 225 −1.670 65.552131.758 1.00 11.21 0 ATOM 2171 CD1 PHE 225 −2.925 65.248 131.239 1.0011.66 0 ATOM 2172 CD2 PHE 225 −1.396 66.869 132.131 1.00 11.41 0 ATOM2173 CE1 PHE 225 −3.899 66.241 131.086 1.00 11.76 0 ATOM 2174 CE2 PHE225 −2.363 67.869 131.983 1.00 14.72 0 ATOM 2175 CZ PHE 225 −3.61867.553 131.458 1.00 16.73 0 ATOM 2176 C PHE 225 1.405 63.392 130.9571.00 6.68 0 ATOM 2177 O PHE 225 1.902 62.850 129.966 1.00 15.31 0 ATOM2178 N THR 226 1.663 63.022 132.203 1.00 2.00 0 ATOM 2180 CA THR 2262.473 61.872 132.509 1.00 2.00 0 ATOM 2181 CB THR 226 3.898 62.226132.853 1.00 13.51 0 ATOM 2182 OG1 THR 226 4.607 61.012 133.094 1.0013.51 0 ATOM 2184 CG2 THR 226 3.961 63.081 134.086 1.00 13.51 0 ATOM2185 C THR 226 1.809 61.217 133.686 1.00 2.00 0 ATOM 2186 O THR 2261.039 61.861 134.394 1.00 13.51 0 ATOM 2187 N PHE 227 2.086 59.938133.883 1.00 2.00 0 ATOM 2189 CA PHE 227 1.466 59.191 134.957 1.00 2.000 ATOM 2190 CB PHE 227 0.206 58.448 134.472 1.00 18.74 0 ATOM 2191 CGPHE 227 0.362 57.793 133.131 1.00 21.51 0 ATOM 2192 CD1 PHE 227 0.71756.453 133.034 1.00 18.55 0 ATOM 2193 CD2 PHE 227 0.188 58.532 133.9541.00 20.45 0 ATOM 2194 CE1 PHE 227 0.902 55.857 131.787 1.00 19.97 0ATOM 2195 CE2 PHE 227 0.371 57.948 130.698 1.00 14.44 0 ATOM 2196 CZ PHE227 0.729 56.612 130.612 1.00 17.09 0 ATOM 2197 C PHE 227 2.427 58.224135.562 1.00 2.00 0 ATOM 2198 O PHE 227 3.282 57.668 134.888 1.00 20.110 ATOM 2199 N GLY 228 2.294 58.066 136.867 1.00 21.90 0 ATOM 2201 CA GLY228 3.143 57.165 137.607 1.00 20.86 0 ATOM 2202 C GLY 228 2.424 55.859137.815 1.00 24.57 0 ATOM 2203 O GLY 228 1.365 55.615 137.223 1.00 2.000 ATOM 2204 N ALA 229 2.974 55.041 138.704 1.00 2.00 0 ATOM 2206 CA ALA229 2.422 53.722 138.971 1.00 2.00 0 ATOM 2207 CB ALA 229 3.372 52.924139.835 1.00 2.00 0 ATOM 2208 C ALA 229 1.055 53.706 139.576 1.00 2.00 0ATOM 2209 O ALA 229 0.359 52.705 139.460 1.00 2.00 0 ATOM 2210 N GLU 2300.666 54.802 140.220 1.00 2.00 0 ATOM 2212 CA GLU 230 −0.648 54.881140.875 1.00 2.00 0 ATOM 2213 CB GLU 230 −0.787 56.193 141.647 1.0066.07 0 ATOM 2214 CG GLU 230 −1.123 56.014 143.111 1.00 71.18 0 ATOM2215 CD GLU 230 −2.376 55.175 143.337 1.00 2.00 0 ATOM 2216 OE1 GLU 230−2.275 53.918 143.199 1.00 2.00 0 ATOM 2217 OE2 GLU 230 −3.448 55.773143.663 1.00 2.00 0 ATOM 2218 C GLU 230 −1.769 54.777 139.862 1.00 2.000 ATOM 2219 O GLU 230 −2.649 53.922 139.966 1.00 60.28 0 ATOM 2220 N VAL231 −1.684 55.652 138.861 1.00 12.41 0 ATOM 2222 CA VAL 231 −2.64955.749 137.772 1.00 9.72 0 ATOM 2223 CB VAL 231 −2.186 56.789 136.7171.00 2.00 0 ATOM 2224 CG1 VAL 231 −3.304 57.063 135.718 1.00 2.00 0 ATOM2225 CG2 VAL 231 −1.728 58.077 137.396 1.00 2.00 0 ATOM 2226 C VAL 231−2.816 54.393 137.102 1.00 11.88 0 ATOM 2227 O VAL 231 −3.937 53.966136.849 1.00 2.00 0 ATOM 2228 N VAL 232 −1.695 53.723 136.832 1.00 2.000 ATOM 2230 CA VAL 232 −1.678 52.397 136.203 1.00 2.00 0 ATOM 2231 CBVAL 232 −0.245 51.817 136.151 1.00 2.00 0 ATOM 2232 CG1 VAL 232 −0.26450.442 135.548 1.00 2.00 0 ATOM 2233 CG2 VAL 232 0.664 52.724 135.3761.00 2.00 0 ATOM 2234 C VAL 232 −2.533 51.413 136.992 1.00 2.00 0 ATOM2235 O VAL 232 −3.449 50.787 136.454 1.00 2.00 0 ATOM 2236 N ALA 233−2.217 51.292 138.275 1.00 38.57 0 ATOM 2238 CA ALA 233 −2.920 50.395139.173 1.00 38.57 0 ATOM 2239 CB ALA 233 −2.297 50.487 140.542 1.0013.20 0 ATOM 2240 C ALA 233 −4.426 50.681 139.245 1.00 38.57 0 ATOM 2241O ALA 233 −5.255 49.763 139.166 1.00 9.05 0 ATOM 2242 N LYS 234 −4.78051.954 139.397 1.00 2.00 0 ATOM 2244 CA LYS 234 −6.177 52.355 139.4771.00 2.00 0 ATOM 2245 CB LYS 234 −6.270 53.843 139.803 1.00 22.96 0 ATOM2246 CG LYS 234 −5.776 54.160 141.205 1.00 31.42 0 ATOM 2247 CD LYS 234−6.667 53.499 142.257 1.00 42.09 0 ATOM 2248 CE LYS 234 −5.916 53.151143.543 1.00 40.07 0 ATOM 2249 NZ LYS 234 −5.104 51.892 143.444 1.0041.59 0 ATOM 2253 C LYS 234 −6.920 52.036 138.183 1.00 2.00 0 ATOM 2254O LYS 234 −7.936 51.319 138.206 1.00 20.45 0 ATOM 2255 N PHE 235 −6.38952.532 137.061 1.00 14.37 0 ATOM 2257 CA PHE 235 −6.973 52.319 135.7331.00 14.37 0 ATOM 2258 CB PHE 235 −6.055 52.880 134.640 1.00 2.00 0 ATOM2259 CG PHE 235 −6.438 52.452 133.249 1.00 2.00 0 ATOM 2260 CD1 PHE 235−7.352 53.199 132.502 1.00 2.00 0 ATOM 2261 CD2 PHE 235 −5.924 51.270132.706 1.00 2.00 0 ATOM 2262 CE1 PHE 235 −7.760 52.777 131.236 1.002.00 0 ATOM 2263 CE2 PHE 235 −6.316 50.834 131.452 1.00 2.00 0 ATOM 2264CZ PHE 235 −7.242 51.588 130.710 1.00 2.00 0 ATOM 2265 C PHE 235 −7.22950.849 135.444 1.00 14.37 0 ATOM 2266 O PHE 235 −8.312 50.471 134.9771.00 2.00 0 ATOM 2267 N LEU 236 −6.217 50.029 135.690 1.00 3.08 0 ATOM2269 CA LEU 236 −6.339 48.606 135.447 1.00 5.72 0 ATOM 2270 CB LEU 236−5.018 47.900 135.731 1.00 2.00 0 ATOM 2271 CG LEU 236 −3.915 48.136134.710 1.00 2.00 0 ATOM 2272 CD1 LEU 236 −2.699 47.332 135.096 1.002.00 0 ATOM 2273 CD2 LEU 236 −4.405 47.737 133.321 1.00 2.00 0 ATOM 2274C LEU 236 −7.452 47.978 136.278 1.00 10.30 0 ATOM 2275 O LEU 236 −8.38947.398 135.712 1.00 2.00 0 ATOM 2276 N HIS 237 −7.368 48.118 137.6061.00 8.44 0 ATOM 2278 CA HIS 237 −8.368 47.540 138.504 1.00 8.44 0 ATOM2279 CB HIS 237 −8.088 47.885 139.980 1.00 38.56 0 ATOM 2280 CG HIS 237−9.141 47.360 140.935 1.00 46.82 0 ATOM 2281 CD2 HIS 237 −10.323 47.916141.329 1.00 46.01 0 ATOM 2282 ND1 HIS 237 −9.035 46.173 141.599 1.0046.3B 0 ATOM 2284 CE1 HIS 237 −10.103 45.989 142.355 1.00 50.85 0 ATOM2285 NE2 HIS 237 −10.900 47.031 142.210 1.00 49.26 0 ATOM 2287 C HIS 237−9.747 48.033 138.138 1.00 8.44 0 ATOM 2288 O HIS 237 −10.672 47.232138.027 1.00 35.43 0 ATOM 2289 N LYS 238 −9.882 49.346 137.947 1.00 2.000 ATOM 2291 CA LYS 238 −11.183 49.923 137.607 1.00 2.00 0 ATOM 2292 CBLYS 238 −11.071 51.424 137.327 1.00 28.00 0 ATOM 2293 CG LYS 238 −12.42752.103 137.104 1.00 29.80 0 ATOM 2294 CD LYS 238 −12.322 53.628 136.8291.00 36.02 0 ATOM 2295 CE LYS 238 −11.917 54.444 136.083 1.00 32.99 0ATOM 2296 NZ LYS 238 −11.833 55.925 137.845 1.00 30.09 0 ATOM 2300 C LYS238 −11.776 49.215 136.403 1.00 2.00 0 ATOM 2301 O LYS 238 −12.99149.205 136.234 1.00 29.21 0 ATOM 2302 N HIS 239 −10.913 46.607 135.5841.00 2.00 0 ATOM 2304 CA HIS 239 −11.340 47.897 134.384 1.00 2.00 0 ATOM2305 CB HIS 239 −10.784 48.603 133.148 1.00 2.00 0 ATOM 2306 CG HIS 239−11.125 50.056 133.091 1.00 2.00 0 ATOM 2307 CD2 HIS 239 −12.282 50.689132.785 1.00 2.00 0 ATOM 2308 ND1 HIS 239 −10.217 51.046 133.397 1.002.00 0 ATOM 2310 CE1 HIS 239 −10.797 52.225 133.284 1.00 2.00 0 ATOM2311 NE2 HIS 239 −12.052 52.036 132.914 1.00 2.00 0 ATOM 2313 C HIS 239−10.946 46.417 134.352 1.00 2.00 0 ATOM 2314 O HIS 239 −10.938 45.798133.295 1.00 2.00 0 ATOM 2315 N ASP 240 −10.642 45.838 135.503 1.0037.96 0 ATOM 2317 CA ASP 240 −10.251 44.433 135.564 1.00 37.83 0 ATOM2318 CB ASP 240 −11.482 43.512 135.737 1.00 39.81 0 ATOM 2319 CG ASP 240−12.676 43.919 134.870 1.00 47.06 0 ATOM 2320 OD1 ASP 240 −12.687 43.571133.666 1.00 50.73 0 ATOM 2321 OD2 ASP 240 −13.610 44.578 135.396 1.0047.58 0 ATOM 2322 C ASP 240 −9.393 43.985 134.385 1.00 37.76 0 ATOM 2323O ASP 240 −9.626 42.933 133.797 1.00 40.81 0 ATOM 2324 N LEU 241 −8.41744.823 134.040 1.00 2.00 0 ATOM 2326 CA LEU 241 −7.462 44.551 132.9551.00 2.00 0 ATOM 2327 CB LEU 241 −7.066 45.858 132.229 1.00 2.00 0 ATOM2328 CG LEU 241 −8.185 46.590 131.429 1.00 2.00 0 ATOM 2329 CD1 LEU 241−7.795 48.047 131.224 1.00 2.00 0 ATOM 2330 CD2 LEU 241 −8.435 45.899130.087 1.00 2.00 0 ATOM 2331 C LEU 241 −6.225 43.925 133.601 1.00 2.000 ATOM 2332 O LEU 241 −6.157 43.832 134.818 1.00 2.00 0 ATOM 2333 N ASP242 −5.235 43.534 132.817 1.00 2.00 0 ATOM 2335 CA ASP 242 −4.046 42.895133.380 1.00 2.00 0 ATOM 2336 CB ASP 242 −3.974 41.455 132.915 1.0025.69 0 ATOM 2337 CG ASP 242 −4.683 40.511 133.822 1.00 12.63 0 ATOM2338 CD1 ASP 242 −5.751 40.853 134.365 1.00 16.91 0 ATOM 2339 OD2 ASP242 −4.149 39.404 133.976 1.00 15.37 0 ATOM 2340 C AEP 242 −2.711 43.506133.033 1.00 2.00 0 ATOM 2341 O ASP 242 −1.702 43.214 133.671 1.00 25.690 ATOM 2342 N LEU 243 −2.689 44.317 131.994 1.00 23.97 0 ATOM 2344 CALEU 243 −1.440 44.899 131.539 1.00 18.84 0 ATOM 2345 CB LEU 243 −0.64443.799 130.822 1.00 2.00 0 ATOM 2346 CG LEU 243 0.698 43.831 130.0971.00 2.00 0 ATOM 2347 CD1 LEU 243 0.410 43.690 128.660 1.00 2.00 0 ATOM2348 CD2 LEU 243 1.526 45.058 130.397 1.00 2.00 0 ATOM 2349 C LEU 243−1.804 46.013 130.590 1.00 19.91 0 ATOM 2350 O LEU 243 −2.960 46.149130.183 1.00 2.00 0 ATOM 2351 N ILE 244 −0.834 46.849 130.283 1.00 24.040 ATOM 2353 CA ILE 244 −1.070 47.919 129.354 1.00 22.40 0 ATOM 2354 CG2ILE 244 −1.164 49.269 130.067 1.00 2.00 0 ATOM 2355 CG2 ILE 244 −1.23550.405 129.026 1.00 2.00 0 ATOM 2356 CG1 ILE 244 −2.395 49.255 130.9941.00 2.00 0 ATOM 2357 CD1 ILE 244 −2.607 50.530 131.803 1.00 2.00 0 ATOM2358 C ILE 244 0.069 47.902 128.369 1.00 25.54 0 ATOM 2359 O ILE 2441.234 47.715 128.755 1.00 2.00 0 ATOM 2360 N CYS 245 −0.286 48.063127.096 1.00 2.00 0 ATOM 2362 CA CYS 245 0.675 48.074 126.019 1.00 2.000 ATOM 2363 CB CYS 245 0.403 46.930 125.077 1.00 7.03 0 ATOM 2364 SG CYS245 1.849 45.913 124.986 1.00 12.49 0 ATOM 2365 C CYS 245 0.668 49.389125.275 1.00 2.00 0 ATOM 2366 O CYS 245 −0.362 49.842 124.792 1.00 8.030 ATOM 2367 N ARG 246 1.829 50.014 125.235 1.00 2.00 0 ATOM 2369 CA ARG246 1.987 51.277 124.562 1.00 2.00 0 ATOM 2370 CB ARG 246 1.763 52.438125.541 1.00 2.00 0 ATOM 2371 CG ARG 246 2.658 52.453 126.783 1.00 2.000 ATOM 2372 CD ARG 246 3.969 53.245 126.623 1.00 2.00 0 ATOM 2373 NE ARG246 3.714 54.656 126.323 1.00 2.00 0 ATOM 2375 CZ ARG 246 4.610 55.622126.519 1.00 2.00 0 ATOM 2376 NH1 ARG 246 5.842 55.334 126.895 1.00 2.000 ATOM 2379 NH2 ARG 246 4.263 56.881 126.289 1.00 2.00 0 ATOM 2382 C ARG246 3.374 51.342 123.938 1.00 2.00 0 ATOM 2383 O ARG 246 4.216 50.471124.168 1.00 2.00 0 ATOM 2384 N ALA 247 3.605 52.345 123.108 1.00 2.00 0ATOM 2386 CA ALA 247 4.906 52.489 122.484 1.00 2.00 0 ATOM 2387 CB ALA247 4.791 52.272 120.975 1.00 61.76 0 ATOM 2388 C ALA 247 5.351 53.902122.819 1.00 2.00 0 ATOM 2389 O ALA 247 5.806 54.172 123.922 1.00 61.760 ATOM 2390 N HIS 248 5.187 54.801 121.870 1.00 2.00 0 ATOM 2392 CA HIS248 5.512 56.193 122.042 1.00 2.00 0 ATOM 2393 C HIS 248 6.959 56.579122.361 1.00 2.00 0 ATOM 2394 O HIS 248 7.507 57.453 121.672 1.00 2.00 0ATOM 2395 CB HIS 248 4.539 56.793 123.054 1.00 2.00 0 ATOM 2396 CG HIS248 4.255 58.246 122.837 1.00 2.00 0 ATOM 2397 ND HIS 248 5.262 59.147122.594 1.00 2.00 0 ATOM 2398 CE1 HIS 248 4.677 60.327 122.535 1.00 2.000 ATOM 2399 CD2 HIS 248 3.074 58.906 122.913 1.00 2.00 0 ATOM 2400 NE2HIS 248 3.359 60.234 122.721 1.00 2.00 0 ATOM 2402 N GLN 249 7.58855.942 123.348 1.00 7.57 0 ATOM 2404 CA GLN 249 8.967 56.283 123.7311.00 7.57 0 ATOM 2405 CB GLN 249 9.010 56.722 125.192 1.00 30.82 0 ATOM2406 CG GLN 249 7.987 57.759 125.566 1.00 32.63 0 ATOM 2407 CD GLN 2498.159 58.253 126.983 1.00 33.33 0 ATOM 2408 OE1 GLN 249 7.871 59.406127.274 1.00 39.63 0 ATOM 2409 NE2 GLN 249 8.640 57.389 127.874 1.0032.74 0 ATOM 2412 C GLN 249 10.052 55.222 123.529 1.00 7.57 0 ATOM 2413O GLN 249 9.884 54.057 123.888 1.00 28.32 0 ATOM 2414 N VAL 250 11.18555.664 122.988 1.00 20.32 0 ATOM 2416 CA VAL 250 12.339 54.805 122.7351.00 20.32 0 ATOM 2417 CB VAL 250 13.571 55.628 122.300 1.00 6.81 0 ATOM2418 CG1 VAL 250 14.641 54.714 121.706 1.00 6.81 0 ATOM 2419 CG2 VAL 25013.161 56.708 121.311 1.00 6.81 0 ATOM 2420 C VAL 250 12.733 54.034123.998 1.00 20.32 0 ATOM 2421 O VAL 250 12.471 54.474 125.120 1.00 6.810 ATOM 2422 N VAL 251 13.372 52.887 123.797 1.00 2.00 0 ATOM 2424 CA VAL251 13.820 52.023 124.871 1.00 2.00 0 ATOM 2425 CB VAL 251 12.655 51.150125.391 1.00 9.99 0 ATOM 2426 CG1 VAL 251 11.778 51.967 126.311 1.009.99 0 ATOM 2427 CG2 VAL 251 11.803 50.661 124.247 1.00 9.99 0 ATOM 2428C VAL 251 14.960 51.167 124.316 1.00 2.00 0 ATOM 2429 O VAL 251 14.79250.470 123.319 1.00 9.99 0 ATOM 2430 N GLU 252 16.118 51.224 124.9721.00 21.55 0 ATOM 2432 CA GLU 252 17.312 50.509 124.519 1.00 19.34 0ATOM 2433 CB GLU 252 18.152 50.605 125.542 1.00 11.04 0 ATOM 2434 CG GLU252 19.819 50.220 124.933 1.00 18.95 0 ATOM 2435 CD GLU 252 20.92649.966 125.962 1.00 20.92 0 ATOM 2436 OE1 GLU 252 21.938 49.303 125.5971.00 17.37 0 ATOM 2437 OE2 GLU 252 20.782 50.422 127.126 1.00 29.06 0ATOM 2438 C GLU 252 17.144 49.055 124.145 1.00 19.22 0 ATOM 2439 O GLU252 17.838 48.571 123.261 1.00 8.64 0 ATOM 2440 N ASP 253 16.240 48.356124.815 1.00 2.00 0 ATOM 2442 CA ASP 253 16.025 46.936 124.544 1.00 2.000 ATOM 2443 CB ASP 253 16.125 46.144 125.850 1.00 78.16 0 ATOM 2444 CGASP 253 17.543 46.106 126.401 1.00 84.05 0 ATOM 2445 OD1 ASP 253 18.06044.995 126.635 1.00 89.15 0 ATOM 2446 OD2 ASP 253 18.148 47.181 126.5981.00 87.62 0 ATOM 2447 C ASP 253 14.710 46.645 123.836 1.00 2.00 0 ATOM2448 O ASP 253 14.201 45.528 123.881 1.00 63.68 0 ATOM 2449 N GLY 25414.161 47.660 123.181 1.00 2.00 0 ATOM 2451 CA GLY 254 12.919 47.483122.457 1.00 2.00 0 ATOM 2452 C GLY 254 11.715 47.547 123.354 1.00 2.000 ATOM 2453 O GLY 254 10.694 48.124 122.993 1.00 43.70 0 ATOM 2454 N TYR255 11.823 46.922 124.516 1.00 2.00 0 ATOM 2456 CA TYR 255 10.749 46.956125.484 1.00 2.00 0 ATOM 2457 CB TYR 255 10.047 45.614 125.571 1.00 2.000 ATOM 2458 CG TYR 255 10.853 44.510 126.185 1.00 2.00 0 ATOM 2459 CD1TYR 255 12.103 44.175 125.696 1.00 2.00 0 ATOM 2460 CE1 TYR 255 12.82443.121 126.248 1.00 2.00 0 ATOM 2461 CD2 TYR 255 10.337 43.770 127.2461.00 2.00 0 ATOM 2462 CE2 TYR 255 11.041 42.721 127.802 1.00 2.00 0 ATOM2463 CZ TYR 255 12.279 42.397 127.301 1.00 2.00 0 ATOM 2464 OH TYR 25512.946 41.328 127.839 1.00 2.00 0 ATOM 2466 C TYR 255 11.262 47.375126.847 1.00 2.00 0 ATOM 2467 O TYR 255 12.453 47.283 127.150 1.00 2.000 ATOM 2468 N GLU 256 10.336 47.853 127.656 1.00 2.00 0 ATOM 2470 CA GLU256 10.632 48.341 128.979 1.00 2.00 0 ATOM 2471 CB GLU 256 11.092 49.798128.910 1.00 20.70 0 ATOM 2472 CG GLU 256 11.264 50.457 130.269 1.0027.98 0 ATOM 2473 CD GLU 256 11.485 51.961 130.179 1.00 31.28 0 ATOM2474 OE1 GLU 256 10.726 52.725 130.834 1.00 33.21 0 ATOM 2475 OE2 GLU256 12.418 52.378 129.453 1.00 39.87 0 ATOM 2476 C GLU 256 9.362 48.238129.800 1.00 2.00 0 ATOM 2477 O GLU 256 8.299 48.743 129.420 1.00 18.740 ATOM 2478 N PHE 257 9.474 47.540 130.919 1.00 2.00 0 ATOM 2480 CA PHE257 8.357 47.364 131.806 1.00 2.00 0 ATOM 2481 CB PHE 257 8.578 46.139132.661 1.00 2.00 0 ATOM 2482 CG PHE 257 8.222 44.865 131.978 1.00 2.000 ATOM 2483 CD1 PHE 257 9.187 43.926 131.694 1.00 2.00 0 ATOM 2484 CD2PHE 257 6.906 44.594 131.649 1.00 2.00 0 ATOM 2485 CE1 PHE 257 8.84942.740 131.101 1.00 2.00 0 ATOM 2486 CE2 PHE 257 6.560 43.417 131.0581.00 2.00 0 ATOM 2487 CZ PHE 257 7.533 42.461 130.781 1.00 2.00 0 ATOM2488 C PHE 257 8.257 48.593 132.671 1.00 2.00 0 ATOM 2489 O PHE 2579.215 49.360 132.782 1.00 2.00 0 ATOM 2490 N PHE 258 7.104 48.777133.286 1.00 25.02 0 ATOM 2492 CA PHE 258 6.884 49.916 134.147 1.0025.02 0 ATOM 2493 CB PHE 258 6.417 51.086 133.299 1.00 2.00 0 ATOM 2494CG PHE 258 5.831 52.237 134.072 1.00 2.00 0 ATOM 2495 CD1 PHE 258 6.62553.325 134.431 1.00 2.00 0 ATOM 2496 CD2 PHE 258 4.468 52.271 134.3691.00 2.00 0 ATOM 2497 CE1 PHE 258 6.070 54.439 135.070 1.00 2.00 0 ATOM2498 CE2 PHE 258 3.900 53.369 135.003 1.00 2.00 0 ATOM 2499 CZ PHE 2584.698 54.461 135.356 1.00 2.00 0 ATOM 2500 C PHE 258 5.837 49.508135.160 1.00 25.02 0 ATOM 2501 O PHE 258 4.965 48.675 134.859 1.00 2.000 ATOM 2502 N ALA 259 5.953 50.063 136.368 1.00 13.61 0 ATOM 2504 CA ALA259 5.027 49.775 137.457 1.00 11.22 0 ATOM 2505 CB ALA 259 3.626 50.318137.122 1.00 2.00 0 ATOM 2506 C ALA 259 4.951 48.286 137.729 1.00 12.730 ATOM 2507 O ALA 259 3.876 47.718 137.660 1.00 2.00 0 ATOM 2508 N LYS260 6.091 47.657 138.016 1.00 2.00 0 ATOM 2510 CA LYS 260 6.140 46.214138.304 1.00 9.39 0 ATOM 2511 CB LYS 260 5.594 45.925 139.704 1.00 20.750 ATOM 2512 CG LYS 260 6.589 46.151 140.844 1.00 28.17 0 ATOM 2513 CDLYS 260 6.797 47.622 141.177 1.00 36.91 0 ATOM 2514 CE LYS 260 7.61547.765 142.449 1.00 36.02 0 ATOM 2515 NZ LYS 260 6.919 47.158 143.6331.00 29.33 0 ATOM 2519 C LYS 260 5.370 45.375 137.279 1.00 2.00 0 ATOM2520 O LYS 260 4.443 44.633 137.641 1.00 15.95 0 ATOM 2521 N ARG 2615.764 45.531 136.004 1.00 2.00 0 ATOM 2523 CA ARG 261 5.173 44.843134.851 1.00 2.00 0 ATOM 2524 CB ARG 261 5.410 43.335 134.964 1.00 21.350 ATOM 2525 CG ARG 261 6.881 42.969 134.928 1.00 21.35 0 ATOM 2526 CDARG 261 7.088 41.473 134.970 1.00 9.03 0 ATOM 2527 NE ARG 261 8.40841.097 134.460 1.00 14.18 0 ATOM 2529 CZ ARG 261 8.801 39.842 134.2221.00 14.38 0 ATOM 2530 NH1 ARG 261 7.976 38.820 134.462 1.00 16.12 0ATOM 2533 NH2 ARG 261 10.018 39.603 133.729 1.00 18.11 0 ATOM 2536 C ARG261 3.692 45.136 134.592 1.00 2.00 0 ATOM 2537 O ARG 261 3.009 44.343133.933 1.00 21.35 0 ATOM 2538 N GLN 262 3.215 46.283 135.092 1.00 7.730 ATOM 2540 CA GLN 262 1.821 46.727 134.927 1.00 7.73 0 ATOM 2541 CB GLN262 1.379 47.620 136.100 1.00 2.00 0 ATOM 2542 CG GLN 262 1.106 46.888137.434 1.00 2.00 0 ATOM 2543 CD GLN 262 0.859 47.840 138.601 1.00 2.000 ATOM 2544 OE1 GLN 262 −0.277 48.137 138.930 1.00 2.00 0 ATOM 2545 NEGLN 262 1.922 48.304 139.237 1.00 2.00 0 ATOM 2548 C GLN 262 1.63547.495 133.628 1.00 7.73 0 ATOM 2549 O GLN 262 0.526 47.650 133.154 1.002.00 0 ATOM 2550 N LEU 263 2.730 47.996 133.078 1.00 2.00 0 ATOM 2552 CALEU 263 2.723 48.726 131.819 1.00 2.00 0 ATOM 2553 CB LEU 263 2.75450.244 132.069 1.00 2.00 0 ATOM 2554 CG LEU 263 3.070 51.201 130.8901.00 2.00 0 ATOM 2555 CD1 LEU 263 2.404 52.532 131.133 1.00 2.00 0 ATOM2556 CD2 LEU 263 4.575 51.408 130.679 1.00 2.00 0 ATOM 2557 C LEU 2633.991 48.305 131.089 1.00 2.00 0 ATOM 2558 O LEU 263 5.018 48.082131.736 1.00 2.00 0 ATOM 2559 N VAL 264 3.932 48.207 129.759 1.00 42.920 ATOM 2561 CA VAL 264 5.105 47.858 128.949 1.00 41.59 0 ATOM 2562 CBVAL 264 5.014 46.373 128.433 1.00 2.00 0 ATOM 2563 CG1 VAL 264 3.95246.242 127.372 1.00 2.00 0 ATOM 2564 CG2 VAL 264 6.349 45.908 127.9291.00 2.00 0 ATOM 2565 C VAL 264 5.193 48.840 127.772 1.00 42.79 0 ATOM2566 O VAL 264 4.164 49.261 127.242 1.00 2.00 0 ATOM 2567 N THR 2656.409 49.237 127.400 1.00 2.00 0 ATOM 2569 CA THR 265 6.639 50.177126.292 1.00 2.00 0 ATOM 2570 CB THR 265 7.420 51.412 126.789 1.00 2.000 ATOM 2571 OG1 THR 265 6.534 52.253 127.531 1.00 2.00 0 ATOM 2573 CG2THR 265 8.021 52.191 125.648 1.00 2.00 0 ATOM 2574 C THR 265 7.40549.523 125.132 1.00 2.00 0 ATOM 2575 O THR 265 8.612 49.245 125.246 1.002.00 0 ATOM 2576 N LEU 266 6.712 49.280 124.019 1.00 7.19 0 ATOM 2578 CALEU 266 7.330 48.653 122.857 1.00 7.19 0 ATOM 2579 CB LEU 266 6.33847.721 122.176 1.00 2.00 0 ATOM 2580 CG LEU 266 5.815 46.505 122.9481.00 2.00 0 ATOM 2581 CD1 LEU 266 4.859 45.733 122.057 1.00 2.00 0 ATOM2582 CD2 LEU 266 6.955 45.599 123.368 1.00 2.00 0 ATOM 2583 C LEU 2667.898 49.618 121.819 1.00 7.19 0 ATOM 2584 O LEU 266 7.329 50.675121.537 1.00 2.00 0 ATOM 2585 N PHE 267 9.033 49.249 121.247 1.00 2.00 0ATOM 2587 CA PHE 267 9.665 50.068 120.222 1.00 2.00 0 ATOM 2588 CB PHE267 10.763 50.960 120.821 1.00 2.00 0 ATOM 2589 CG PHE 267 10.937 52.260120.091 1.00 2.00 0 ATOM 2590 CD1 PHE 267 9.985 53.261 120.206 1.00 2.000 ATOM 2591 CD2 PHE 267 12.021 52.468 119.255 1.00 2.00 0 ATOM 2592 CE1PHE 267 10.108 54.445 119.493 1.00 2.00 0 ATOM 2593 CE2 PHE 267 12.14653.652 118.540 1.00 2.00 0 ATOM 2594 CZ PHE 267 11.187 54.636 118.6611.00 2.00 0 ATOM 2595 C PHE 267 10.246 49.076 119.227 1.00 2.00 0 ATOM2596 O PHE 267 11.418 48.693 119.321 1.00 2.00 0 ATOM 2597 N SER 2689.387 48.638 118.302 1.00 12.94 0 ATOM 2599 CA SER 268 9.707 47.648117.267 1.00 12.94 0 ATOM 2600 CB SER 268 8.420 47.043 116.723 1.0011.11 0 ATOM 2601 OG SER 268 7.593 46.579 117.771 1.00 11.11 0 ATOM 2603C SER 268 10.450 48.231 116.103 1.00 12.94 0 ATOM 2604 O SER 268 10.15049.344 115.711 1.00 11.11 0 ATOM 2605 N ALA 269 11.391 47.454 115.5561.00 71.01 0 ATOM 2607 CA ALA 269 12.222 47.779 114.378 1.00 71.72 0ATOM 2608 CB ALA 269 11.383 48.513 113.295 1.00 82.00 0 ATOM 2609 C ALA269 13.585 48.469 114.543 1.00 74.82 0 ATOM 2610 O ALA 269 14.609 47.889114.186 1.00 91.21 0 ATOM 2611 N PRO 270 13.619 49.704 115.076 1.0028.82 0 ATOM 2612 CD PRO 270 12.436 50.466 115.513 1.00 2.00 0 ATOM 2613CA PRO 270 14.788 50.542 115.316 1.00 31.86 0 ATOM 2614 CB PRO 27014.340 51.374 116.502 1.00 2.00 0 ATOM 2615 CG PRO 270 13.034 51.783116.023 1.00 2.00 0 ATOM 2616 C PRO 270 16.254 50.192 115.428 1.00 29.520 ATOM 2617 O PRO 270 16.716 49.072 115.272 1.00 2.00 0 ATOM 2618 N ASN271 16.928 51.318 115.593 1.00 2.00 0 ATOM 2620 CA ASN 271 18.346 51.621115.754 1.00 2.00 0 ATOM 2621 CB ASN 271 19.168 50.935 114.663 1.0035.96 0 ATOM 2622 CG ASN 271 18.483 50.953 113.300 1.00 61.96 0 ATOM2623 OD1 ASN 271 17.605 51.787 113.034 1.00 36.11 0 ATOM 2624 ND2 ASN271 18.872 50.015 112.432 1.00 36.31 0 ATOM 2627 C ASN 271 18.199 53.143115.475 1.00 2.00 0 ATOM 2628 O ASN 271 19.072 53.807 114.915 1.00 61.690 ATOM 2629 N TYR 272 17.039 53.638 115.930 1.00 2.00 0 ATOM 2631 CA TYR272 16.489 54.978 115.793 1.00 2.00 0 ATOM 2632 CB TYR 272 15.772 55.358117.090 1.00 17.33 0 ATOM 2633 CG TYR 272 14.528 56.208 116.897 1.0010.41 0 ATOM 2634 CD1 TYR 272 13.458 55.769 116.105 1.00 11.86 0 ATOM2635 CE1 TYR 272 12.313 56.571 115.917 1.00 14.32 0 ATOM 2636 CD2 TYR272 14.421 57.454 117.492 1.00 10.66 0 ATOM 2637 CE2 TYR 272 13.28758.256 117.312 1.00 15.43 0 ATOM 2638 CZ TYR 272 12.243 57.813 116.5291.00 9.40 0 ATOM 2639 OH TYR 272 11.143 58.636 116.375 1.00 17.38 0 ATOM2641 C TYR 272 17.332 56.132 115.299 1.00 2.03 0 ATOM 2642 O TYR 27218.356 56.471 115.888 1.00 29.84 0 ATOM 2643 N CYS 273 16.860 56.731114.201 1.00 7.24 0 ATOM 2645 CA CYS 273 17.490 57.875 113.553 1.00 6.820 ATOM 2646 CB CYS 273 17.201 59.167 114.331 1.00 11.61 0 ATOM 2647 SGCYS 273 15.496 59.459 114.843 1.00 10.42 0 ATOM 2648 C CYS 273 19.01057.724 113.411 1.00 6.43 0 ATOM 2649 O CYS 273 19.738 58.717 113.5121.00 9.04 0 ATOM 2650 N GLY 274 19.498 56.502 113.185 1.00 13.82 0 ATOM2652 CA GLY 274 20.934 56.303 113.044 1.00 13.82 0 ATOM 2653 C GLY 27421.710 57.029 114.134 1.00 13.82 0 ATOM 2654 O GLY 274 22.817 57.515113.906 1.00 39.65 0 ATOM 2655 N GLU 275 21.120 57.085 115.326 1.0092.80 0 ATOM 2657 CA GLU 275 21.719 57.764 116.467 1.00 91.32 0 ATOM2658 CB GLU 275 21.479 59.276 116.340 1.00 25.01 0 ATOM 2659 CG GLU 27519.994 59.656 116.224 1.00 31.82 0 ATOM 2660 CD GLU 275 19.730 61.158116.060 1.00 30.92 0 ATOM 2661 OE1 GLU 275 18.951 61.713 116.868 1.0038.31 0 ATOM 2662 OE2 GLU 275 20.276 61.787 115.125 1.00 34.18 0 ATOM2663 C GLU 275 21.137 57.260 117.794 1.00 90.32 0 ATOM 2664 O GLU 27520.941 58.047 118.716 1.00 23.69 0 ATOM 2665 N PHE 276 20.868 55.958117.901 1.00 16.78 0 ATOM 2667 CA PHE 276 20.303 55.412 119.134 1.0017.07 0 ATOM 2668 CB PHE 276 18.774 55.551 119.125 1.00 37.20 0 ATOM2669 CG PHE 276 18.280 56.876 119.652 1.00 35.89 0 ATOM 2670 CD1 PHE 27617.522 57.726 118.849 1.00 35.53 0 ATOM 2671 CD2 PHE 276 18.572 57.273120.949 1.00 39.17 0 ATOM 2672 CE1 PHE 276 17.067 58.945 119.329 1.0031.45 0 ATOM 2673 CE2 PHE 276 18.123 58.485 121.434 1.00 36.03 0 ATOM2674 CZ PHE 276 17.367 59.324 120.621 1.00 39.75 0 ATOM 2675 C PHE 27620.681 53.992 119.597 1.00 17.79 0 ATOM 2676 O PHE 276 20.463 53.669120.772 1.00 36.38 0 ATOM 2677 N ASP 277 21.213 53.140 118.712 1.0027.69 0 ATOM 2679 CA ASP 277 21.638 51.772 119.093 1.00 27.89 0 ATOM2680 CB ASP 277 22.884 51.848 120.016 1.00 0.27 0 ATOM 2681 CG ASP 27723.231 50.508 120.716 1.00 39.20 0 ATOM 2682 OD1 ASP 277 23.163 50.470121.964 1.00 39.20 0 ATOM 2683 OD2 ASP 277 23.576 49.508 120.036 1.0039.20 0 ATOM 2684 C ASP 277 20.560 50.892 119.742 1.00 26.41 0 ATOM 2685O ASP 277 20.761 49.675 119.918 1.00 0.76 0 ATOM 2686 N ASN 278 19.42851.491 120.101 1.00 2.00 0 ATOM 2688 CA ASN 278 18.344 50.758 120.7221.00 2.00 0 ATOM 2689 CB ASN 278 17.142 51.676 120.920 1.00 2.00 0 ATOM2690 CG ASN 278 16.394 51.930 119.639 1.00 2.00 0 ATOM 2691 OD1 ASN 27816.867 52.643 118.765 1.00 2.00 0 ATOM 2692 ND2 ASN 278 15.225 51.324119.510 1.00 2.00 0 ATOM 2695 C ASN 278 17.938 49.602 119.829 1.00 2.000 ATOM 2696 O ASN 278 17.970 49.710 118.600 1.00 2.00 0 ATOM 2697 N ALA279 17.603 48.487 120.454 1.00 14.31 0 ATOM 2699 CA ALA 279 17.13347.320 119.734 1.00 14.31 0 ATOM 2700 CB ALA 279 17.281 46.081 120.5941.00 2.00 0 ATOM 2701 C ALA 279 15.655 47.631 119.498 1.00 14.31 0 ATOM2702 O ALA 279 15.155 48.645 119.996 1.00 2.00 0 ATOM 2703 N GLY 28014.959 46.786 118.743 1.00 2.00 0 ATOM 2705 CA GLY 280 13.549 47.014118.484 1.00 2.00 0 ATOM 2706 C GLY 280 12.864 45.783 118.992 1.00 2.000 ATOM 2707 O GLY 280 13.381 44.698 118.799 1.00 9.35 0 ATOM 2708 N ALA281 11.725 45.906 119.643 1.00 2.00 0 ATOM 2710 CA ALA 281 11.082 44.702120.167 1.00 2.00 0 ATOM 2711 CB ALA 281 10.799 44.850 121.658 1.00 2.000 ATOM 2712 C ALA 281 9.815 44.345 119.437 1.00 2.00 0 ATOM 2713 O ALA281 9.451 44.992 118.473 1.00 2.00 0 ATOM 2714 N MET 282 9.140 43.312119.918 1.00 14.19 0 ATOM 2716 CA MET 282 7.907 42.833 119.315 1.0014.19 0 ATOM 2717 CB MET 282 8.232 42.115 117.995 1.00 25.98 0 ATOM 2718CG MET 282 7.056 41.885 117.060 1.00 24.91 0 ATOM 2719 SD MET 282 7.30440.449 115.980 1.00 25.45 0 ATOM 2720 CE MET 282 9.044 40.644 115.5251.00 21.20 0 ATOM 2721 C MET 282 7.397 41.851 120.363 1.00 14.19 0 ATOM2722 O MET 282 8.142 40.972 120.790 1.00 24.98 0 ATOM 2723 N MET 2836.148 42.004 120.788 1.00 2.00 0 ATOM 2725 CA MET 283 5.592 41.143121.825 1.00 2.00 0 ATOM 2726 CB MET 283 4.940 41.991 122.929 1.00 19.290 ATOM 2727 CG MET 283 4.481 41.205 124.142 1.00 19.76 0 ATOM 2728 SDMET 283 3.228 42.077 125.116 1.00 20.92 0 ATOM 2729 CE MET 283 4.20443.235 125.966 1.00 17.38 0 ATOM 2730 C MET 283 4.592 40.125 121.3391.00 2.00 0 ATOM 2731 O MET 283 3.456 40.470 121.036 1.00 16.17 0 ATOM2732 N SER 284 5.012 35.571 121.291 1.00 2.00 0 ATOM 2734 CA SER 2844.148 37.777 120.869 1.00 2.00 0 ATOM 2735 CB SER 284 4.991 36.664120.220 1.00 20.90 0 ATOM 2736 OG SER 284 6.376 36.771 120.528 1.0023.67 0 ATOM 2738 C SBR 284 3.275 37.202 122.018 1.00 2.00 0 ATOM 2739 OSER 284 3.777 36.595 122.978 1.00 26.24 0 ATOM 2740 N VAL 285 1.96737.402 121.894 1.00 2.00 0 ATOM 2742 CA VAL 285 0.979 36.940 122.8591.00 2.00 0 ATOM 2743 CB VAL 285 −0.091 38.026 123.056 1.00 2.00 0 ATOM2744 CG1 VAL 285 −0.952 37.714 124.262 1.00 2.00 0 ATOM 2745 CG2 VAL 2850.572 39.381 123.171 1.00 2.00 0 ATOM 2746 C VAL 285 0.274 35.644122.417 1.00 2.00 0 ATGK 2747 O VAL 285 −0.572 35.672 121.532 1.00 2.000 ATOM 2748 N ASP 286 0.609 34.512 123.026 1.00 2.00 0 ATOM 2750 CA ASP286 −0.056 33.258 122.663 1.00 2.00 0 ATOM 2751 CB ASP 286 0.771 32.019123.071 1.00 22.05 0 ATOM 2752 CG ASP 286 1.192 32.020 124.531 1.0024.93 0 ATOM 2753 OD1 ASP 286 0.578 32.742 125.344 1.00 32.49 0 ATOM2754 OD2 ASP 286 2.152 31.285 124.866 1.00 28.80 0 ATOM 2755 C ASP 286−1.494 33.141 123.181 1.00 2.00 0 ATOM 2756 O ASP 286 −1.993 34.023123.881 1.00 12.72 0 ATOM 2757 N GLU 287 −2.144 32.036 122.817 1.0028.94 0 ATOM 2759 CA GLU 287 −3.543 31.744 123.153 1.00 28.07 0 ATOM2760 CB GLU 287 −3.912 30.340 122.657 1.00 59.96 0 ATOM 2761 CG GLU 287−3.610 30.049 121.177 1.00 66.13 0 ATOM 2762 CD GLU 287 −2.139 29.730120.890 1.00 70.85 0 ATOM 2763 OE1 GLU 287 −1.712 29.890 119.723 1.0075.38 0 ATOM 2764 OE2 GLU 287 −1.412 29.315 121.822 1.00 73.00 0 ATOM2765 C GLU 287 −3.876 31.836 124.638 1.00 30.16 0 ATOM 2766 O GLU 287−5.017 32.074 125.029 1.00 62.37 0 ATOM 2767 N THR 288 −2.852 31.654125.455 1.00 41.83 0 ATOM 2769 CA THR 288 −2.995 31.665 126.894 1.0036.69 0 ATOM 2770 CB THR 288 −2.269 30.449 127.457 1.00 11.26 0 ATOM2771 OG1 THR 288 −0.865 30.552 127.171 1.00 11.65 0 ATOM 2773 CG2 THR288 −2.827 29.171 126.800 1.00 11.00 0 ATOM 2774 C THR 288 −2.461 32.947127.533 1.00 36.73 0 ATOM 2775 O THR 288 −2.015 32.948 128.675 1.0018.83 0 ATOM 2776 N LEU 289 −2.492 34.034 126.776 1.00 6.34 0 ATOM 2778CA LEU 289 −2.027 35.334 127.232 1.00 2.00 0 ATOM 2779 CB LEU 289 −3.08135.946 125.155 1.00 2.00 0 ATOM 2780 CG LEU 289 −4.282 36.465 127.3461.00 2.00 0 ATOM 2751 CD1 LEU 289 −5.390 36.973 128.263 1.00 2.00 0 ATOM2782 CD2 LEU 289 −3.817 37.593 126.412 1.00 2.00 0 ATOM 2783 C LEU 289−0.615 35.454 127.826 1.00 2.00 0 ATOM 2784 O LEU 289 −0.299 36.445128.493 1.00 2.00 0 ATOM 2785 N MET 290 0.244 34.475 127.566 1.00 20.850 ATOM 2787 CA MET 290 1.619 34.548 128.046 1.00 23.75 0 ATOM 2788 CBMET 290 2.238 33.156 128.212 1.00 24.81 0 ATOM 2789 CG NET 290 3.71733.200 128.623 1.00 23.46 0 ATOM 2790 SD MET 290 4.726 31.867 127.9351.00 26.75 0 ATOM 2791 CE MET 290 5.196 32.555 126.310 1.00 29.16 0 ATOM2792 C MET 290 2.430 35.332 127.015 1.00 19.20 0 ATOM 2793 O MET 2902.545 34.910 125.858 1.00 29.26 0 ATOM 2794 N CYS 291 3.003 36.454127.449 1.00 2.00 0 ATOM 2796 CA CYS 291 3.797 37.336 126.590 1.00 2.000 ATOM 2797 CB CYS 291 1.624 38.773 127.062 1.00 17.87 0 ATOM 2798 SGCYS 291 1.913 39.123 127.404 1.00 13.14 0 ATOM 2799 C CYS 291 5.29337.034 126.465 1.00 2.00 0 ATOM 2800 O CYS 291 5.958 36.694 127.448 1.0023.09 0 ATOM 2801 N SER 292 5.808 37.187 125.244 1.00 5.71 0 ATOM 2803CA SER 292 7.223 36.977 124.928 1.00 9.30 0 ATOM 2804 CB SER 292 7.38535.709 124.098 1.00 12.57 0 ATOM 2805 OG SER 292 6.548 34.675 124.5931.00 20.05 0 ATOM 2807 C SER 292 7.691 38.195 124.113 1.00 4.04 0 ATOM2808 O SER 292 6.867 38.867 123.485 1.00 14.24 0 ATOM 2809 N PHE 2938.993 38.487 124.128 1.00 2.00 0 ATOM 2811 CA PHE 293 9.526 39.623123.385 1.00 2.00 0 ATOM 2812 CB PHE 293 10.077 40.676 124.337 1.0017.01 0 ATOM 2813 CG PHE 293 9.063 41.244 125.270 1.00 19.48 0 ATOM 2814CD1 PHE 293 8.772 40.611 126.466 1.00 23.05 0 ATOM 2815 CD2 PHE 2938.414 42.426 124.967 1.00 21.44 0 ATOM 2816 CE1 PHE 293 7.852 41.146127.352 1.00 24.82 0 ATOM 2817 CE2 PHE 293 7.494 42.968 125.845 1.0020.24 0 ATOM 2818 CZ PHE 293 7.213 42.325 127.044 1.00 22.28 0 ATOM 2819C PHE 293 10.641 39.259 122.412 1.00 2.00 0 ATOM 2820 O PHE 293 11.71538.839 122.853 1.00 17.14 0 ATOM 2821 N GLN 294 10.390 39.418 121.1041.00 2.00 0 ATOM 2823 CA GLN 294 11.404 39.162 120.060 1.00 2.00 0 ATOM2824 CB GLN 294 10.748 38.742 118.743 1.00 14.58 0 ATOM 2825 CG GLN 29410.074 37.383 116.798 1.00 22.67 0 ATOM 2826 CD GLN 294 8.684 37.387116.179 1.00 24.62 0 ATOM 2827 OE1 GLN 294 7.835 38.206 116.535 1.0027.17 0 ATOM 2828 NE2 GLN 294 8.441 36.456 117.258 1.00 25.84 0 ATOM2831 C GLN 294 12.182 40.464 119.855 1.00 2.00 0 ATOM 2832 O GLN 29411.587 41.528 119.660 1.00 20.63 0 ATOM 2833 N ILE 295 13.507 40.379119.893 1.00 11.23 0 ATOM 2835 CA ILE 295 14.356 41.569 119.772 1.0012.32 0 ATOM 2836 CB ILE 295 15.400 41.624 120.968 1.00 10.45 0 ATOM2837 CG2 ILE 295 16.277 42.875 120.886 1.00 10.45 0 ATOM 2838 CG1 ILE295 14.674 41.607 122.324 1.00 10.45 0 ATOM 2839 CD1 ILE 295 13.75642.807 122.580 1.00 10.45 0 ATOM 2840 C ILE 295 15.126 41.703 118.4531.00 7.79 0 ATOM 2841 O ILE 295 15.723 40.741 117.974 1.00 10.45 0 ATOM2842 N LEU 296 15.087 42.900 117.873 1.00 11.71 0 ATOM 2844 CA LEU 29615.824 43.235 116.658 1.00 13.33 0 ATOM 2845 CB LEU 296 14.987 44.102115.717 1.00 24.46 0 ATOM 2846 CG LEU 296 13.835 43.467 114.946 1.0022.89 0 ATOM 2847 CD1 LEU 296 12.626 43.340 115.862 1.00 22.12 0 ATOM2848 CD2 LEU 296 13.499 44.331 113.732 1.00 19.71 0 ATOM 2849 C LEU 29616.965 44.070 117.224 1.00 13.82 0 ATOM 2850 O LEU 296 16.785 45.260117.485 1.00 13.73 0 ATOM 2851 N LYS 297 18.135 43.454 117.381 1.00 2.000 ATOM 2853 CA LYS 297 19.319 44.087 117.997 1.00 2.00 0 ATOM 2854 CBLYS 297 20.224 42.971 118.527 1.00 79.20 0 ATOM 2855 CG LYS 297 20.79242.094 117.419 1.00 79.65 0 ATOM 2856 CD LYS 297 21.898 41.187 117.9151.00 76.76 0 ATOM 2857 CE LYS 297 22.543 40.430 116.762 1.00 71.01 0ATOM 2858 NZ LYS 297 23.644 39.543 117.232 1.00 66.76 0 ATOM 2862 C LYS297 20.259 45.149 117.376 1.00 2.00 0 ATOM 2863 O LYS 297 21.255 45.501118.013 1.00 94.11 0 ATOM 2864 N PRO 298 19.995 45.656 116.152 1.0040.11 0 ATOM 2865 CD PRO 298 18.942 45.297 115.188 1.00 2.00 0 ATOM 2866CA PRO 298 20.900 46.658 115.553 1.00 40.11 0 ATOM 2667 CB PRO 29820.220 46.981 114.227 1.00 2.00 0 ATOM 2868 CG PRO 298 19.570 45.700113.886 1.00 2.00 0 ATOM 2869 C PRO 298 21.264 47.935 116.312 1.00 40.110 ATOM 2870 O PRO 298 21.041 48.056 117.514 1.00 2.00 0 ATOM 2871 N ALA299 21.854 48.878 115.574 1.00 61.74 0 ATOM 2873 CA ALA 299 22.27650.176 116.101 1.00 61.74 0 ATOM 2874 CB ALA 299 23.627 50.063 116.7991.00 2.00 0 ATOM 2875 C ALA 299 22.373 51.193 114.973 1.00 61.74 0 ATOM2876 O ALA 299 21.893 52.319 115.105 1.00 2.00 0 ATOM 2677 N ASN 50841.191 29.848 91.500 1.00 48.97 0 ATOM 2879 CA ASN 508 39.902 30.15090.896 1.00 48.97 0 ATOM 2880 CB ASN 508 38.951 30.831 91.887 1.00 0.740 ATOM 2881 CG ASN 508 37.666 31.359 91.203 1.00 0.74 0 ATOM 2882 OD1ASN 508 36.879 30.587 90.592 1.00 0.74 0 ATOM 2883 ND2 ASN 508 37.45332.682 91.290 1.00 0.74 0 ATOM 2886 C ASN 508 40.096 31.041 89.687 1.0048.97 0 ATOM 2887 O ASN 508 40.274 32.266 89.798 1.00 0.74 0 ATOM 2888 NILE 509 40.006 30.387 88.533 1.00 17.27 0 ATOM 2890 CA ILE 509 40.18830.983 87.224 1.00 18.75 0 ATOM 2891 CB ILE 509 40.088 29.889 86.1271.00 44.37 0 ATOM 2892 CG2 ILE 509 38.650 29.700 85.673 1.00 52.59 0ATOM 2893 CG1 ILE 509 41.013 30.238 84.966 1.00 44.31 0 ATOM 2894 CD1ILE 509 42.489 30.093 85.305 1.00 44.07 0 ATOM 2895 C ILE 509 39.19632.089 86.957 1.00 18.83 0 ATOM 2896 O ILE 509 39.463 32.956 86.131 1.0044.94 0 ATOM 2897 N ASP 510 38.070 32.071 87.670 1.00 2.00 0 ATOM 2899CA ASP 510 37.048 33.088 87.485 1.00 2.00 0 ATOM 2900 CB ASP 510 35.71132.605 88.031 1.00 52.76 0 ATOM 2901 CG ASP 510 35.029 31.634 87.0891.00 54.58 0 ATOM 2902 OD1 ASP 510 34.008 32.015 86.483 1.00 54.54 0ATOM 2903 OD2 ASP 510 35.521 30.497 86.944 1.00 58.16 0 ATOM 2904 C ASP510 37.403 34.442 88.056 1.00 2.00 0 ATOM 2905 O ASP 510 37.016 35.46487.490 1.00 52.34 0 ATOM 2906 N SER 511 38.143 34.471 89.160 1.00 2.00 0ATOM 2908 CA SER 511 38.548 35.763 89.739 1.00 2.00 0 ATOM 2909 CB SER511 39.372 35.570 91.015 1.00 53.95 0 ATOM 2910 OG SER 511 38.631 34.90792.020 1.00 50.17 0 ATOM 2912 C SER 511 39.447 36.379 88.690 1.00 2.00 0ATOM 2913 O SER 511 39.228 37.488 88.203 1.00 59.62 0 ATOM 2914 N ILE512 40.453 35.584 88.349 1.00 45.56 0 ATOM 2916 CA ILE 512 41.462 35.89287.361 1.00 40.32 0 ATOM 2917 CB ILE 512 42.224 34.598 87.035 1.00 2.000 ATOM 2918 CG2 ILE 512 43.250 34.842 85.943 1.00 2.00 0 ATOM 2919 CG1ILE 512 42.897 34.092 88.316 1.00 2.00 0 ATOM 2920 CD1 ILE 512 43.63532.772 88.174 1.00 2.00 0 ATOM 2921 C ILE 512 40.843 36.514 86.105 1.0041.22 0 ATOM 2922 O ILE 512 41.051 37.700 85.848 1.00 2.00 0 ATOM 2923 NILE 513 40.061 35.728 85.358 1.00 2.00 0 ATOM 2925 CA ILE 513 39.40836.195 84.136 1.00 2.00 0 ATOM 2926 CB ILE 513 38.435 35.123 83.529 1.002.00 0 ATOM 2927 CG2 ILE 513 37.560 35.757 82.455 1.00 2.00 0 ATOM 2928CG1 ILE 513 39.227 33.963 82.890 1.00 2.00 0 ATOM 2929 CD1 ILE 51338.365 32.796 82.367 1.00 2.00 0 ATOM 2930 C ILE 513 38.640 37.49384.335 1.00 2.00 0 ATOM 2931 O ILE 513 38.764 38.403 83.519 1.00 2.00 0ATOM 2932 N GLN 514 37.858 37.611 85.402 1.00 12.46 0 ATOM 2934 CA GLN514 37.117 38.853 85.581 1.00 12.46 0 ATOM 2935 CB GLN 514 35.985 38.67886.591 1.00 26.62 0 ATOM 2936 CG GLN 514 36.384 38.251 87.972 1.00 26.620 ATOM 2937 CD GLN 514 35.195 38.279 88.929 1.00 26.62 0 ATOM 2938 OE1GLN 514 34.582 39.337 89.140 1.00 26.62 0 ATOM 2939 NE2 GLN 514 34.85137.116 89.502 1.00 26.62 0 ATOM 2942 C GLN 514 38.009 40.055 85.936 1.0012.46 0 ATOM 2943 O GLN 514 37.686 41.205 85.613 1.00 26.62 0 ATOM 2944N ARG 515 39.147 39.773 86.564 1.00 6.50 0 ATOM 2946 CA ARG 515 40.09040.810 86.937 1.00 7.53 0 ATOM 2947 CB ARG 515 41.125 40.251 87.917 1.008.04 0 ATOM 2948 CG ARG 515 40.626 40.257 89.362 1.00 8.04 0 ATOM 2949CD ARG 515 41.182 39.117 90.212 1.00 8.04 0 ATOM 2950 NE ARG 515 42.60939.250 90.490 1.00 8.04 0 ATOM 2952 CZ ARG 515 43.355 38.276 90.998 1.008.04 0 ATOM 2953 NH1 ARG 515 42.505 37.103 91.275 1.00 8.04 0 ATOM 2956NH2 ARG 515 44.646 38.478 91.228 1.00 8.04 0 ATOM 2959 C ARG 515 40.74541.325 85.669 1.00 12.92 0 ATOM 2960 O ARG 515 40.840 42.537 85.464 1.008.04 0 ATOM 2961 N LEU 516 41.167 40.398 84.810 1.00 8.93 0 ATOM 2963 CALEU 516 41.788 40.727 83.525 1.00 6.71 0 ATOM 2964 CB LEU 516 42.17239.431 82.786 1.00 2.00 0 ATOM 2965 CG LEU 516 43.298 38.561 83.385 1.002.00 0 ATOM 2966 CD1 LEU 516 43.057 37.096 83.082 1.00 2.00 0 ATOM 2967CD2 LEU 516 44.650 38.986 82.843 1.00 2.00 0 ATOM 2968 C LEU 516 40.80941.562 82.675 1.00 5.82 0 ATOM 2969 O LEU 516 41.187 42.516 82.002 1.002.00 0 ATOM 2970 N LEU 517 39.534 41.228 82.755 1.00 2.00 0 ATOM 2972 CALEU 517 38.519 41.928 81.993 1.00 2.73 0 ATOM 2973 CB LEU 517 37.33640.993 81.752 1.00 7.08 0 ATOM 2974 CG LEU 517 37.203 40.297 80.398 1.003.18 0 ATOM 2975 CD1 LEU 517 38.524 39.694 79.950 1.00 12.86 0 ATOM 2976CD2 LEU 517 36.133 39.227 80.518 1.00 9.89 0 ATOM 2977 C LEU 517 37.98943.262 82.536 1.00 9.79 0 ATOM 2978 O LEU 517 37.169 43.908 81.858 1.0016.46 0 ATOM 2979 N GLU 518 38.409 43.708 83.722 1.00 2.00 0 ATOM 2981CA GLU 518 37.845 44.981 84.178 1.00 2.00 0 ATOM 2982 CB GLU 518 37.62145.015 85.699 1.00 2.00 0 ATOM 2983 CG GLU 518 36.908 46.315 86.231 1.002.00 0 ATOM 2964 CD GLU 518 35.550 46.705 85.526 1.00 2.00 0 ATOM 2985OE1 GLU 518 34.554 47.001 86.262 1.00 2.00 0 ATOM 2986 OE2 GLU 51835.481 46.748 84.252 1.00 2.00 0 ATOM 2987 C GLU 518 38.618 46.20083.677 1.00 2.00 0 ATOM 2988 O GLU 518 38.121 47.335 83.729 1.00 2.00 0ATOM 2989 N VAL 519 39.828 45.978 83.166 1.00 21.94 0 ATOM 2991 CA VAL519 40.582 47.094 82.589 1.00 19.66 0 ATOM 2992 CB VAL 519 42.078 46.80482.428 1.00 40.73 0 ATOM 2993 CG1 VAL 519 42.759 46.806 83.771 1.0042.53 0 ATOM 2994 CG2 VAL 519 42.272 45.488 81.684 1.00 37.62 0 ATOM2995 C VAL 519 40.007 47.307 81.196 1.00 22.58 0 ATOM 2996 O VAL 51940.402 48.229 80.493 1.00 48.07 0 ATOM 2997 N ARG 520 39.066 46.44280.815 1.00 26.87 0 ATOM 2999 CA ARG 520 38.410 46.495 79.515 1.00 28.580 ATOM 3000 CB ARG 520 37.216 45.534 79.480 1.00 40.18 0 ATOM 3001 CGARG 520 36.878 45.043 78.086 1.00 44.14 0 ATOM 3002 CD ARG 520 35.99443.814 78.116 1.00 47.95 0 ATOM 3003 NE ARG 520 34.592 44.131 77.8721.00 44.40 0 ATOM 3005 CZ ARG 520 33.565 43.402 78.308 1.00 51.11 0 ATOM3006 NH1 ARG 520 33.767 42.298 79.022 1.00 48.24 0 ATOM 3009 NH2 ARG 52032.325 43.778 78.026 1.00 49.03 0 ATOM 3012 C ARG 520 37.946 47.89979.147 1.00 28.36 0 ATOM 3013 O ARG 520 37.881 48.247 77.964 1.00 46.630 ATOM 3014 N GLY 521 37.639 48.703 80.158 1.00 27.79 0 ATOM 3016 CA GLY521 37.186 50.056 79.905 1.00 30.97 0 ATOM 3017 C GLY 521 38.158 51.09980.400 1.00 32.39 0 ATOM 3018 O GLY 521 37.739 52.124 80.940 1.00 54.550 ATOM 3019 N SER 522 39.451 50.840 80.225 1.00 46.95 0 ATOM 3021 CA SER522 40.495 51.761 80.663 1.00 47.91 0 ATOM 3022 CB SER 522 41.300 51.14681.801 1.00 33.41 0 ATOM 3023 OG SER 522 40.459 50.694 82.844 1.00 32.380 ATOM 3025 C SER 522 41.436 52.094 79.517 1.00 49.38 0 ATOM 3026 O SER522 41.495 51.378 78.513 1.00 34.02 0 ATOM 3027 N LYS 523 42.174 53.18479.679 1.00 84.47 0 ATOM 3029 CA LYS 523 43.127 53.640 78.674 1.00 81.240 ATOM 3030 CB LYS 523 44.035 54.715 79.274 1.00 43.78 0 ATOM 3031 CGLYS 523 43.307 55.987 79.619 1.00 44.40 0 ATOM 3032 CD LYS 523 44.20656.990 80.322 1.00 68.84 0 ATOM 3033 CE LYS 523 43.493 58.336 80.4581.00 44.38 0 ATOM 3034 NZ LYS 523 42.092 58.186 80.983 1.00 44.06 0 ATOM3038 C LYS 523 44.000 52.501 78.173 1.00 80.03 0 ATOM 3039 O LYS 52344.314 51.584 78.937 1.00 43.77 0 ATOM 3040 N PRO 524 44.360 52.51476.873 1.00 2.00 0 ATOM 3041 CD PRO 524 43.901 53.427 75.811 1.00 84.060 ATOM 3042 CA PRO 524 45.218 51.459 76.316 1.00 2.00 0 ATOM 3043 CB PRO524 45.357 51.871 74.850 1.00 62.26 0 ATOM 3044 CG PRO 524 44.057 52.57474.576 1.00 78.74 0 ATOM 3045 C PRO 524 46.576 51.489 77.063 1.00 2.00 0ATOM 3046 O PRO 524 47.510 52.190 76.653 1.00 76.15 0 ATOM 3047 N GLY525 46.647 50.742 78.166 1.00 33.80 0 ATOM 3049 CA GLY 525 47.846 50.68278.984 1.00 98.68 0 ATOM 3050 C GLY 525 47.575 50.197 80.403 1.00 99.000 ATOM 3051 O GLY 525 48.427 49.555 81.013 1.00 2.00 0 ATOM 3052 N LYS526 46.397 50.496 80.942 1.00 49.37 0 ATOM 3054 CA LYS 526 46.047 50.07082.297 1.00 44.35 0 ATOM 3055 CB LYS 526 44.537 50.226 82.505 1.00 0.790 ATOM 3056 CG LYS 526 44.013 49.851 83.911 1.00 0.82 0 ATOM 3057 CD LYS526 44.370 50.915 84.950 1.00 0.29 0 ATOM 3058 CE LYS 526 44.413 50.31286.354 1.00 28.48 0 ATOM 3059 NZ LYS 526 45.166 51.143 87.348 1.00 28.160 ATOM 3063 C LYS 526 46.461 48.602 82.479 1.00 41.81 0 ATOM 3064 O LYS526 45.984 47.727 81.760 1.00 0.75 0 ATOM 3065 N ASN 527 47.372 46.33783.412 1.00 6.60 0 ATOM 3067 CA ASN 527 47.861 46.972 83.639 1.00 6.60 0ATOM 3068 CB ASN 527 49.208 46.977 84.361 1.00 20.38 0 ATOM 3069 CG ASN527 50.300 47.733 83.645 1.00 30.42 0 ATOM 3070 OD1 ASN 527 50.69548.820 84.068 1.00 29.99 0 ATOM 3071 ND2 ASN 527 50.818 47.148 82.5611.00 30.17 0 ATOM 3074 C ASN 527 46.895 46.135 84.463 1.00 6.60 0 ATOM3075 O ASN 527 45.866 46.628 84.924 1.00 23.47 0 ATOM 3076 N VAL S2847.271 44.866 84.647 1.00 26.67 0 ATOM 3078 CA VAL 528 46.541 43.87485.443 1.00 26.67 0 ATOM 3079 CB VAL 528 45.575 43.004 84.603 1.00 33.440 ATOM 3080 CG1 VAL 528 44.909 41.956 85.488 1.00 33.01 0 ATOM 3081 CG2VAL 528 44.524 43.858 83.956 1.00 35.61 0 ATOM 3082 C VAL 528 47.60742.940 86.018 1.00 26.67 0 ATOM 3083 O VAL 528 48.054 42.003 85.348 1.0036.85 0 ATOM 3084 N GLN 529 48.048 43.231 87.239 1.00 2.00 0 ATOM 3086CA GLN 529 49.061 42.407 87.920 1.00 2.00 0 ATOM 3087 CB GLN 529 49.96843.297 88.787 1.00 30.41 0 ATOM 3088 CG GLN 529 51.142 42.569 89.4281.00 32.34 0 ATOM 3089 CD GLN 529 52.380 42.558 86.558 1.00 31.37 0 ATOM3090 OE1 GLN 529 53.436 43.035 88.962 1.00 35.47 0 ATOM 3091 NE2 GLN 52952.259 42.014 87.361 1.00 30.69 0 ATOM 3094 C GLN 529 48.350 41.34788.792 1.00 2.00 0 ATOM 3095 O GLN 529 47.545 41.689 89.663 1.00 32.29 0ATOM 3096 N LEU 530 48.624 40.072 88.536 1.00 2.00 0 ATOM 3098 CA LEU530 47.994 39.002 89.303 1.00 2.00 0 ATOM 3099 CB LEU 530 47.462 37.91188.381 1.00 12.99 0 ATOM 3100 CG LEU 530 46.535 38.302 87.248 1.00 12.990 ATOM 3101 CD1 LEU 530 47.297 38.159 85.956 1.00 12.99 0 ATOM 3102 CD2LEU 530 45.313 37.403 87.243 1.00 12.99 0 ATOM 3103 C LEU 530 49.00238.365 90.236 1.00 2.00 0 ATOM 3104 O LEU 530 50.207 38.543 90.067 1.0012.99 0 ATOM 3105 N GLN 531 48.516 37.622 91.221 1.00 2.00 0 ATOM 3107CA GLN 531 49.418 36.932 92.117 1.00 2.00 0 ATOM 3108 CB GLN 531 46.63436.051 93.090 1.00 61.54 0 ATOM 3109 CG GLN 531 48.376 36.639 94.4651.00 62.91 0 ATOM 3110 CD GLN 531 47.221 37.603 94.487 1.00 63.71 0 ATOM3111 OE1 GLN 531 47.322 36.684 95.053 1.00 59.60 0 ATOM 3112 NE2 GLN 53146.111 37.219 93.876 1.00 61.96 0 ATOM 3115 C GLN 531 50.337 36.04091.266 1.00 2.00 0 ATOM 3116 O GLN 531 49.859 35.276 90.422 1.00 67.15 0ATOM 3117 N GLU 532 51.647 36.153 91.480 1.00 2.00 0 ATOM 3119 CA GLU532 52.655 35.349 90.766 1.00 2.00 0 ATOM 3120 CB GLU 532 54.056 35.62391.336 1.00 19.01 0 ATOM 3121 CG GLU 532 55.176 34.623 90.966 1.00 22.740 ATOM 3122 CD GLU 532 56.496 34.871 91.746 1.00 25.50 0 ATOM 3123 OE1GLU 532 56.845 36.042 92.058 1.00 20.16 0 ATOM 3124 OE2 GLU 532 57.19533.879 92.051 1.00 20.82 0 ATOM 3125 C GLU 532 52.345 33.866 90.900 1.002.00 0 ATOM 3126 O GLU 532 52.833 33.059 90.116 1.00 17.28 0 ATOM 3127 NASN 533 51.568 33.509 91.918 1.00 26.78 0 ATOM 3129 CA ASN 533 51.18132.120 92.127 1.00 27.69 0 ATOM 3130 CB ASN 533 50.751 31.857 93.5871.00 42.65 0 ATOM 3131 CG ASN 533 49.834 32.944 94.154 1.00 49.17 0 ATOM3132 OD1 ASN 533 50.274 34.070 94.392 1.00 50.06 0 ATOM 3133 ND2 ASN 53348.568 32.605 94.387 1.00 50.92 0 ATOM 3136 C ASN 533 50.053 31.77091.165 1.00 26.24 0 ATOM 3137 O ASN 533 49.918 30.615 90.747 1.00 41.800 ATOM 3138 N GLU 534 49.253 32.777 90.815 1.00 24.76 0 ATOM 3140 CA GLU534 48.146 32.592 89.893 1.00 19.83 0 ATOM 3141 CB GLU 534 47.228 33.80189.916 1.00 17.15 0 ATOM 3142 CG GLU 534 46.509 33.924 91.239 1.00 23.750 ATOM 3143 CD GLU 534 45.598 35.132 91.321 1.00 25.22 0 ATOM 3144 OE1GLU 534 44.490 34.997 91.890 1.00 24.06 0 ATOM 3145 OE2 GLU 534 45.99136.215 90.822 1.00 29.17 0 ATOM 3146 C GLU 534 48.712 32.365 88.510 1.0021.16 0 ATOM 3147 O GLU 534 48.235 31.500 87.779 1.00 14.12 0 ATOM 3148N ILE 535 49.747 33.122 88.156 1.00 2.00 0 ATOM 3150 CA ILE 535 50.38932.942 86.859 1.00 2.00 0 ATOM 3151 CB ILE 535 51.442 34.023 86.570 1.002.00 0 ATOM 3152 CG2 ILE 535 52.050 33.784 85.190 1.00 2.00 0 ATOM 3153CG1 ILE 535 50.793 35.411 86.641 1.00 2.00 0 ATOM 3154 CD1 ILE 53551.537 36.488 85.849 1.00 2.00 0 ATOM 3155 C ILE 535 51.060 31.56586.797 1.00 2.00 0 ATOM 3156 O ILE 535 50.788 30.780 85.876 1.00 2.00 0ATOM 3157 N ARG 536 51.914 31.256 87.778 1.00 2.00 0 ATOM 3159 CA ARG536 52.583 29.951 87.793 1.00 2.00 0 ATOM 3160 CB ARG 536 53.495 29.81889.011 1.00 39.13 0 ATOM 3161 CG ARG 536 52.808 29.604 90.341 1.00 45.430 ATOM 3162 CD ARG 536 53.839 29.648 91.474 1.00 49.26 0 ATOM 3163 NEARG 536 55.059 28.910 91.132 1.00 56.04 0 ATOM 3165 CZ ARG 536 55.11027.603 90.876 1.00 55.24 0 ATOM 3166 NH1 ARG 536 54.011 26.860 90.9201.00 53.92 0 ATOM 3169 NH2 ARG 536 56.267 27.032 90.576 1.00 44.45 0ATOM 3172 C ARG 536 51.562 28.811 87.742 1.00 2.00 0 ATOM 3173 O ARG 53651.806 27.778 87.126 1.00 37.01 0 ATOM 3174 N GLY 537 50.406 29.03388.363 1.00 2.00 0 ATOM 3176 CA GLY 537 49.345 28.046 88.340 1.00 2.00 0ATOM 3177 C GLY 537 48.690 28.000 86.970 1.00 2.00 0 ATOM 3178 O GLY 53748.292 26.924 86.512 1.00 2.00 0 ATOM 3179 N LEU 538 48.574 29.16486.319 1.00 9.51 0 ATOM 3181 CA LEU 538 47.979 29.276 84.977 1.00 7.63 0ATOM 3182 CB LEU 538 47.930 30.736 84.513 1.00 2.00 0 ATOM 3183 CG LEU538 46.654 31.524 84.772 1.00 2.00 0 ATOM 3184 CD1 LEU 538 46.865 32.96684.408 1.00 2.00 0 ATOM 3185 CD2 LEU 538 45.531 30.940 83.972 1.00 2.000 ATOM 3186 C LEU 538 48.816 28.472 84.000 1.00 12.73 0 ATOM 3187 O LEU538 48.295 27.635 83.271 1.00 2.00 0 ATOM 3188 N CYS 539 50.120 28.72484.000 1.00 2.00 0 ATOM 3190 CA CYS 539 51.036 28.001 83.132 1.00 2.00 0ATOM 3191 CB CYS 539 52.473 28.494 83.337 1.00 20.52 0 ATOM 3192 SC CYS539 52.713 30.280 83.567 1.00 19.48 0 ATOM 3193 C CYS 539 50.957 26.50183.474 1.00 2.00 0 ATOM 3194 O CYS 539 50.854 25.658 82.588 1.00 29.69 0ATOM 3195 N LEU 540 50.984 26.191 84.769 1.00 26.60 0 ATOM 3197 CA LEU540 50.919 24.819 85.265 1.00 26.60 0 ATOM 3198 CB LEU 540 51.106 24.81886.786 1.00 2.00 0 ATOM 3199 CG LEU 540 52.539 24.770 87.339 1.00 2.00 0ATOM 3200 CD1 LEU 540 53.001 23.360 87.385 1.00 2.00 0 ATOM 3201 CD2 LEU540 53.492 25.577 86.497 1.00 2.00 0 ATOM 3202 C LEU 540 49.635 24.06184.899 1.00 26.60 0 ATOM 3203 O LEU 540 49.677 22.856 84.619 1.00 2.00 0ATOM 3204 N LYS 541 48.500 24.756 84.901 1.00 13.81 0 ATOM 3206 CA LYS541 47.219 24.126 84.561 1.00 17.46 0 ATOM 3207 CB LYS 541 46.046 24.96465.108 1.00 14.61 0 ATOM 3208 CG LYS 541 45.844 24.880 86.624 1.00 22.530 ATOM 3209 CD LYS 541 44.709 25.777 87.150 1.00 31.64 0 ATOM 3210 CELYS 541 45.175 27.201 87.517 1.00 37.15 0 ATOM 3211 NZ LYS 541 44.14728.010 88.284 1.00 27.95 0 ATOM 3215 C LYS 541 47.047 23.891 83.046 1.0016.97 0 ATOM 3216 O LYS 541 46.862 22.745 82.608 1.00 8.92 0 ATOM 3217 NSER 542 47.131 24.963 82.253 1.00 2.00 0 ATOM 3219 CA SER 542 46.97524.877 80.791 1.00 2.00 0 ATOM 3220 CB SER 542 47.165 26.256 80.150 1.002.00 0 ATOM 3221 OG SER 542 48.499 26.703 80.298 1.00 2.00 0 ATOM 3223 CSER 542 47.915 23.870 80.107 1.00 2.00 0 ATOM 3224 O SER 542 47.45022.991 79.377 1.00 2.00 0 ATOM 3225 N ARG 543 49.223 24.016 80.338 1.008.64 0 ATOM 3227 CA ARG 543 50.244 23.128 79.772 1.00 8.64 0 ATOM 3228CB ARG 543 51.607 23.381 80.434 1.00 2.00 0 ATOM 3229 CG ARG 543 52.67622.339 80.116 1.00 4.46 0 ATOM 3230 CD ARG 543 53.757 22.313 81.173 1.002.00 0 ATOM 3231 NE ARG 543 54.989 21.663 80.708 1.00 2.00 0 ATOM 3233CZ ARG 543 55.326 20.395 80.952 1.00 2.00 0 ATOM 3234 NH1 ARG 543 54.52319.606 81.656 1.00 3.99 0 ATOM 3237 NH2 ARG 543 56.483 19.915 80.5081.00 2.00 0 ATOM 3240 C ARG 543 49.873 21.660 79.981 1.00 8.64 0 ATOM3241 O ARG 543 50.187 20.806 79.146 1.00 5.52 0 ATOM 3242 N GLU 54449.227 21.352 81.098 1.00 42.94 0 ATOM 3244 CA GLU 544 48.834 19.97881.355 1.00 44.22 0 ATOM 3245 CB GLU 544 48.308 19.841 82.778 1.00 37.010 ATOM 3246 CG GLU 544 48.175 18.415 83.249 1.00 41.48 0 ATOM 3247 CDGLU 544 47.561 18.341 84.626 1.00 46.37 0 ATOM 3248 OE1 GLU 544 46.43318.858 84.791 1.00 53.09 0 ATOM 3249 OE2 GLU 544 48.202 17.777 85.5431.00 47.19 0 ATOM 3250 C GLU 544 47.763 19.584 80.339 1.00 40.49 0 ATOM3251 O GLU 544 47.742 18.450 79.867 1.00 34.80 0 ATOM 3252 N ILE 54546.898 20.538 79.994 1.00 2.00 0 ATOM 3254 CA ILE 545 45.823 20.31579.017 1.00 2.00 0 ATOM 3255 CB ILE 545 44.805 21.487 78.959 1.00 11.870 ATOM 3256 CG2 ILE 545 43.645 21.102 78.064 1.00 6.53 0 ATOM 3257 CG1ILE 545 44.288 21.836 80.353 1.00 16.65 0 ATOM 3258 CD1 ILE 545 43.26722.955 80.361 1.00 13.01 0 ATOM 3259 C ILE 545 46.377 20.149 77.605 1.002.00 0 ATOM 3260 O ILE 545 45.960 19.267 76.868 1.00 9.05 0 ATOM 3261 NPHE 546 47.302 21.018 77.227 1.00 2.00 0 ATOM 3263 CA PHE 546 47.89520.945 75.918 1.00 2.00 0 ATOM 3264 CB PHE 546 49.058 21.906 75.816 1.002.00 0 ATOM 3265 CG PHE 546 48.653 23.335 75.875 1.00 2.00 0 ATOM 3266CD1 PHE 546 49.512 24.293 76.399 1.00 2.00 0 ATOM 3267 CD2 PHE 54647.411 23.730 75.426 1.00 2.00 0 ATOM 3268 CE1 PHE 546 49.127 25.63076.473 1.00 2.00 0 ATOM 3269 CE2 PHE 546 47.023 25.050 75.496 1.00 2.000 ATOM 3270 CZ PHE 546 47.881 26.006 76.020 1.00 2.00 0 ATOM 3271 C PHE546 48.374 19.538 75.658 1.00 2.00 0 ATOM 3272 O PHE 546 48.141 18.99074.596 1.00 2.00 0 ATOM 3273 N LEU 547 49.012 18.935 76.647 1.00 2.00 0ATOM 3275 CA LEU 547 49.527 17.582 76.506 1.00 2.00 0 ATOM 3276 CB LEU547 50.499 17.289 77.654 1.00 6.91 0 ATOM 3277 CG LEU 547 51.754 18.15977.582 1.00 8.40 0 ATOM 3278 CD1 LEU 547 52.096 18.701 78.934 1.00 16.170 ATOM 3279 CD2 LEU 547 52.905 17.363 77.029 1.00 11.72 0 ATOM 3280 CLEU 547 48.422 16.526 76.434 1.00 2.00 0 ATOM 3281 O LEU 547 48.64215.414 75.946 1.00 6.91 0 ATOM 3282 N SER 548 47.230 16.881 76.903 1.005.34 0 ATOM 3284 CA SER 548 46.091 15.964 76.898 1.00 5.34 0 ATOM 3285CB SER 548 45.099 16.356 77.993 1.00 36.19 0 ATOM 3286 OG SER 548 45.61117.389 78.821 1.00 39.64 0 ATOM 3288 C SER 548 45.374 15.975 75.552 1.005.34 0 ATOM 3289 O SER 548 44.299 15.381 75.406 1.00 36.66 0 ATOM 3290 NGLN 549 45.968 16.649 74.569 1.00 23.79 0 ATOM 3292 CA GLN 549 45.38416.754 73.242 1.00 23.94 0 ATOM 3293 CB GLN 549 44.751 18.128 73.0841.00 46.80 0 ATOM 3294 CG GLN 549 43.567 18.354 73.990 1.00 38.43 0 ATOM3295 CD GLN 549 43.180 19.800 74.054 1.00 38.53 0 ATOM 3296 OE1 GLN 54943.934 20.672 73.624 1.00 46.42 0 ATOM 3297 NE2 GLN 549 42.002 20.07474.595 1.00 39.54 0 ATOM 3300 C GLN 549 46.456 16.547 72.182 1.00 25.870 ATOM 3301 O GLN 549 47.634 16.766 72.451 1.00 42.01 0 ATOM 3302 N PRO550 46.068 16.105 70.963 1.00 2.00 0 ATOM 3303 CD PRO 550 44.729 15.73870.469 1.00 36.68 0 ATOM 3304 CA PRO 550 47.075 15.894 69.921 1.00 2.000 ATOM 3305 CB PRO 550 46.253 15.350 68.750 1.00 36.68 0 ATOM 3306 CGPRO 550 44.886 15.915 68.988 1.00 36.68 0 ATOM 3307 C PRO 550 47.81617.184 69.580 1.00 2.00 0 ATOM 3308 O PRO 550 47.318 18.280 69.840 1.0036.68 0 ATOM 3309 N ILE 551 49.013 17.046 69.015 1.00 2.00 0 ATOM 3311CA ILE 551 49.824 18.198 68.626 1.00 2.00 0 ATOM 3312 CB ILE 551 51.29317.788 68.510 1.00 2.00 0 ATOM 3313 CG2 ILE 551 51.490 16.914 67.2921.00 2.00 0 ATOM 3314 CG1 ILE 551 52.182 19.021 68.475 1.00 2.00 0 ATOM3315 CD1 ILE 551 53.648 18.701 68.468 1.00 2.00 0 ATOM 3316 C ILE 55149.295 18.705 67.281 1.00 2.00 0 ATOM 3317 O ILE 551 49.596 19.80766.840 1.00 2.00 0 ATOM 3318 N LEU 552 48.514 17.863 66.627 1.00 2.00 0ATOM 3320 CA LEU 552 47.896 18.202 65.369 1.00 2.00 0 ATOM 3321 CB LEU552 48.287 17.148 64.320 1.00 2.00 0 ATOM 3322 CG LEU 552 47.809 17.21562.867 1.00 2.00 0 ATOM 3323 CD1 LEU 552 48.082 15.532 62.209 1.00 2.000 ATOM 3324 CD2 LEU 552 48.544 16.174 62.131 1.00 2.00 0 ATOM 3325 C LEU552 46.397 18.180 65.728 1.00 2.00 0 ATOM 3326 O LEU 552 45.753 17.12465.734 1.00 2.00 0 ATOM 3327 N LEU 553 45.881 19.352 66.100 1.00 20.15 0ATOM 3329 CA LEU 553 44.485 19.512 66.517 1.00 20.15 0 ATOM 3330 CB LEU553 44.240 20.926 67.055 1.00 2.00 0 ATOM 3331 CG LEU 553 44.374 21.26268.543 1.00 2.00 0 ATOM 3332 CD1 LEU 553 45.224 20.282 69.290 1.00 2.000 ATOM 3333 CD2 LEU 553 44.942 22.639 68.638 1.00 2.00 0 ATOM 3334 C LEU553 43.495 19.229 65.410 1.00 20.15 0 ATOM 3335 O LEU 553 43.604 19.78064.309 1.00 2.00 0 ATOM 3336 N GLU 554 42.524 18.375 65.724 1.00 14.62 0ATOM 3338 CA GLU 554 41.476 17.983 64.786 1.00 13.80 0 ATOM 3339 CB GLU554 41.135 16.492 64.956 1.00 57.31 0 ATOM 3340 CG GLU 554 42.326 15.55065.177 1.00 69.83 0 ATOM 3341 CD GLU 554 43.168 15.310 63.927 1.00 73.580 ATOM 3342 OE1 GLU 554 42.918 15.957 62.887 1.00 82.45 0 ATOM 3343 OE2GLU 554 44.090 14.465 63.988 1.00 79.54 0 ATOM 3344 C GLU 554 40.24118.823 65.105 1.00 12.33 0 ATOM 3345 O GLU 554 39.253 18.311 65.638 1.0051.99 0 ATOM 3346 N LEU 555 40.296 20.112 64.792 1.00 2.00 0 ATOM 3348CA LEU 555 39.176 21.007 65.077 1.00 2.00 0 ATOM 3349 CB LEU 555 39.62822.456 64.911 1.00 2.00 0 ATOM 3350 CG LEU 555 40.660 22.840 65.972 1.002.00 0 ATOM 3351 CD1 LEU 555 41.274 24.194 65.697 1.00 2.00 0 ATOM 3352CD2 LEU 555 39.959 22.839 67.294 1.00 2.00 0 ATOM 3353 C LEU 555 37.93220.734 64.237 1.00 2.00 0 ATOM 3354 O LEU 555 37.862 19.731 63.537 1.002.00 0 ATOM 3355 N GLU 556 36.933 21.603 64.345 1.00 2.00 0 ATOM 3357 CAGLU 556 35.716 21.460 63.569 1.00 2.00 0 ATOM 3358 CB GLU 556 35.02820.115 63.846 1.00 32.90 0 ATOM 3359 CG GLU 556 34.175 20.063 65.1001.00 48.42 0 ATOM 3360 CD GLU 556 32.935 19.176 64.941 1.00 58.49 0 ATOM3361 OE1 GLU 556 32.792 18.188 65.711 1.00 60.59 0 ATOM 3362 OE2 GLU 55632.101 19.477 64.047 1.00 59.64 0 ATOM 3363 C GLU 556 34.736 22.58363.826 1.00 2.00 0 ATOM 3364 O GLU 556 35.067 23.580 64.446 1.00 16.83 0ATOM 3365 N ALA 557 33.544 22.423 63.257 1.00 2.00 0 ATOM 3367 CA ALA557 32.398 23.330 63.427 1.00 2.00 0 ATOM 3368 CB ALA 557 31.747 23.01364.788 1.00 47.62 0 ATOM 3369 C ALA 557 32.701 24.826 63.286 1.00 2.00 0ATOM 3370 O ALA 557 33.711 25.191 62.671 1.00 40.85 0 ATOM 3371 N PRO558 31.811 25.711 63.802 1.00 2.00 0 ATOM 3372 CD PRO 558 30.459 25.55864.353 1.00 2.44 0 ATOM 3373 CA PRO 558 32.123 27.139 63.658 1.00 2.00 0ATOM 3374 CB PRO 558 30.786 27.826 63.951 1.00 2.53 0 ATOM 3375 CG PRO558 29.798 26.759 63.843 1.00 2.44 0 ATOM 3376 C PRO 558 33.190 27.64864.627 1.00 2.00 0 ATOM 3377 O PRO 558 32.975 27.616 65.844 1.00 7.53 0ATOM 3378 N LEU 559 34.325 28.119 64.112 1.00 2.00 0 ATOM 3380 CA LEU559 35.349 28.689 64.982 1.00 2.00 0 ATOM 3381 CB LEU 559 36.568 27.76965.148 1.00 2.00 0 ATOM 3382 CG LEU 559 37.592 27.571 64.050 1.00 2.00 0ATOM 3383 CD1 LEU 559 38.620 26.563 64.476 1.00 2.00 0 ATOM 3384 CD2 LEU559 36.885 27.083 62.830 1.00 2.00 0 ATOM 3385 C LEU 559 35.769 30.03864.435 1.00 2.00 0 ATOM 3386 O LEU 559 35.444 30.389 63.311 1.00 2.00 0ATOM 3387 N LYS 560 36.461 30.804 65.260 1.00 2.00 0 ATOM 3389 CA LYS560 36.932 32.115 64.889 1.00 2.00 0 ATOM 3390 CB LYS 560 36.359 33.14365.843 1.00 20.44 0 ATOM 3391 CG LYS 560 36.612 34.562 65.427 1.00 20.440 ATOM 3392 CD LYS 560 35.574 35.512 66.018 1.00 20.44 0 ATOM 3393 CELYS 560 34.253 35.452 65.275 1.00 20.44 0 ATOM 3394 NZ LYS 560 33.70634.083 65.225 1.00 20.44 0 ATOM 3398 C LYS 560 38.426 31.990 65.052 1.002.00 0 ATOM 3399 O LYS 560 38.908 31.624 66.117 1.00 20.44 0 ATOM 3400 NILE 561 39.164 32.244 63.986 1.00 2.00 0 ATOM 3402 CA ILE 561 40.60632.106 64.017 1.00 2.00 0 ATOM 3403 CB ILE 561 41.095 31.318 62.750 1.002.00 0 ATOM 3404 CG2 ILE 561 42.566 30.913 62.891 1.00 2.00 0 ATOM 3405CG1 ILE 561 40.204 30.076 62.558 1.00 2.00 0 ATOM 3406 CD1 ILE 56140.806 28.940 61.761 1.00 2.00 0 ATOM 3407 C ILE 561 41.247 33.48864.102 1.00 2.00 0 ATOM 3408 O ILE 561 40.703 34.450 63.543 1.00 2.00 0ATOM 3409 N CYS 562 42.365 33.586 64.833 1.00 2.00 0 ATOM 3411 CA CYS562 43.124 34.829 65.001 1.00 2.00 0 ATOM 3412 CB CYS 562 42.793 35.46466.339 1.00 13.31 0 ATOM 3413 SG CYS 562 41.073 35.733 66.574 1.00 13.310 ATOM 3414 C CYS 562 44.642 34.599 64.928 1.00 2.00 0 ATOM 3415 O CYS562 45.151 33.551 65.373 1.00 13.31 0 ATOM 3416 N GLY 563 45.362 35.54064.348 1.00 2.00 0 ATOM 3418 CA GLY 563 46.805 35.424 64.252 1.00 2.00 0ATOM 3419 C GLY 563 47.546 36.260 65.291 1.00 2.00 0 ATOM 3420 O GLY 56346.997 36.556 66.351 1.00 5.66 0 ATOM 3421 N ASP 564 48.780 36.64964.958 1.00 2.00 0 ATOM 3423 CA ASP 564 49.657 37.452 65.811 1.00 2.00 0ATOM 3424 CB ASP 564 50.705 38.191 64.964 1.00 2.00 0 ATOM 3425 CG ASP564 51.724 37.270 64.321 1.00 2.00 0 ATOM 3426 OD1 ASP 564 51.390 36.56563.334 1.00 2.00 0 ATOM 3427 OD2 ASP 564 52.876 37.249 64.786 1.00 2.000 ATOM 3428 C ASP 564 48.981 38.497 66.693 1.00 2.00 0 ATOM 3429 O ASP564 48.395 39.442 66.189 1.00 2.00 0 ATOM 3430 N ILE 565 49.094 38.32768.009 1.00 31.53 0 ATOM 3432 CA ILE 565 48.537 39.294 68.953 1.00 35.540 ATOM 3433 CB ILE 565 47.810 38.612 70.135 1.00 9.39 0 ATOM 3434 CG2ILE 565 47.139 39.664 70.996 1.00 9.39 0 ATOM 3435 CG1 ILE 565 46.71037.685 69.622 1.00 9.39 0 ATOM 3436 CD1 ILE 565 45.611 38.424 68.9061.00 9.39 0 ATOM 3437 C ILE 565 49.681 40.168 69.489 1.00 32.16 0 ATOM3438 O ILE 565 49.491 41.356 69.756 1.00 9.39 0 ATOM 3439 N HIS 56650.659 39.560 69.637 1.00 14.33 0 ATOM 3441 CA HIS 566 52.082 40.21970.111 1.00 15.67 0 ATOM 3442 C HIS 566 51.958 41.227 71.245 1.00 16.540 ATOM 3443 O HIS 566 52.427 42.359 71.127 1.00 15.89 0 ATOM 3444 CB HIS566 52.811 40.882 68.943 1.00 9.53 0 ATOM 3445 CG HIS 566 53.652 39.93668.153 1.00 9.53 0 ATOM 3446 ND1 HIS 566 54.829 39.391 68.605 1.00 9.530 ATOM 3448 CD2 HIS 566 53.470 39.432 66.910 1.00 9.53 0 ATOM 3449 NE2HIS 566 54.525 38.582 66.589 1.00 9.53 0 ATOM 3450 CE1 HIS 566 55.31238.600 67.652 1.00 9.53 0 ATOM 3451 N GLY 567 51.337 40.808 72.345 1.002.00 0 ATOM 3453 CA GLY 567 51.170 41.686 73.490 1.00 2.00 0 ATOM 3454 CGLY 567 50.249 42.894 73.375 1.00 2.00 0 ATOM 3455 O GLY 567 50.26943.764 74.254 1.00 3.59 0 ATOM 3456 N GLN 568 49.455 42.973 72.314 1.0036.66 0 ATOM 3458 CA GLN 568 48.527 44.086 72.141 1.00 34.60 0 ATOM 3459CB GLN 568 48.164 44.236 70.667 1.00 2.50 0 ATOM 3460 CG GLN 568 49.34544.164 69.715 1.00 3.37 0 ATOM 3461 CD GLN 568 49.768 45.507 69.144 1.003.10 0 ATOM 3462 OE1 GLN 568 50.958 45.769 68.974 1.00 4.87 0 ATOM 3463NE2 GLN 568 48.800 46.349 68.833 1.00 18.91 0 ATOM 3466 C GLN 568 47.28043.730 72.950 1.00 34.26 0 ATOM 3467 O GLN 568 46.253 43.365 72.380 1.007.33 0 ATOM 3468 N TYR 569 47.363 43.864 74.272 1.00 2.00 0 ATOM 3470 CATYR 569 46.263 43.485 75.161 1.00 2.00 0 ATOM 3471 CB TYR 569 46.66443.687 76.620 1.00 26.93 0 ATOM 3472 CG TYR 569 45.692 43.048 77.6001.00 27.02 0 ATOM 3473 CD1 TYR 569 45.401 41.684 77.520 1.00 22.43 0ATOM 3474 CE1 TYR 569 44.496 41.097 78.393 1.00 24.12 0 ATOM 3475 CD2TYR 569 45.049 43.807 78.575 1.00 24.68 0 ATOM 3476 CE2 TYR 569 44.13943.223 79.449 1.00 24.01 0 ATOM 3477 CZ TYR 569 43.866 41.869 79.3521.00 28.35 0 ATOM 3478 OH TYR 569 42.950 41.282 80.197 1.00 35.43 0 ATOM3480 C TYR 569 44.888 44.085 74.946 1.00 2.00 0 ATOM 3481 O TYR 56943.885 43.374 75.015 1.00 29.07 0 ATOM 3482 N TYR 570 44.817 45.38474.710 1.00 11.04 0 ATOM 3484 CA TYR 570 43.515 45.999 74.510 1.00 14.250 ATOM 3485 CB TYR 570 43.599 47.505 74.766 1.00 46.12 0 ATOM 3486 CGTYR 570 43.577 47.792 76.261 1.00 51.43 0 ATOM 3487 CD1 TYR 570 44.74947.743 77.025 1.00 48.11 0 ATOM 3488 CE1 TYR 570 44.715 47.952 78.3981.00 52.04 0 ATOM 3489 CD2 TYR 570 42.375 48.063 76.917 1.00 49.50 0ATOM 3490 CE2 TYR 570 42.340 48.272 78.277 1.00 51.48 0 ATOM 3491 CZ TYR570 43.507 48.215 79.010 1.00 53.24 0 ATOM 3492 OH TYR 570 43.452 48.42680.359 1.00 54.97 0 ATOM 3494 C TYR 570 42.929 45.6S3 73.152 1.00 13.580 ATOM 3495 O TYR 570 41.708 45.654 72.967 1.00 43.14 0 ATOM 3496 N ASP571 43.813 45.311 72.217 1.00 12.28 0 ATOM 3498 CA ASP 571 43.402 44.90370.891 1.00 11.58 0 ATOM 3499 CB ASP 571 44.590 44.870 69.975 1.00 6.850 ATOM 3500 CG ASP 571 45.128 46.237 69.733 1.00 12.88 0 ATOM 3501 OD1ASP 571 46.254 46.522 70.185 1.00 19.21 0 ATOM 3502 OD2 ASP 571 44.40247.044 69.110 1.00 16.01 0 ATOM 3503 C ASP 571 42.818 43.539 71.062 1.0014.93 0 ATOM 3504 O ASP 571 41.775 43.247 70.507 1.00 19.59 0 ATOM 3505N LEU 572 43.487 42.707 71.853 1.00 2.00 0 ATOM 3507 CA LEU 572 42.97741.378 72.147 1.00 2.00 0 ATOM 3508 CB LEU 572 43.909 40.641 73.110 1.002.00 0 ATOM 3509 CG LEU 572 43.302 39.416 73.820 1.00 2.00 0 ATOM 3510CD1 LEU 572 43.061 38.267 72.832 1.00 2.00 0 ATOM 3511 CD2 LEU 57244.227 38.982 74.955 1.00 2.00 0 ATOM 3512 C LEU 572 41.579 41.55472.771 1.00 2.00 0 ATOM 3513 O LEU 572 40.678 40.758 72.512 1.00 2.00 0ATOM 3514 N LEU 573 41.395 42.604 73.572 1.00 9.95 0 ATOM 3516 CA LEU573 40.092 42.874 74.195 1.00 6.25 0 ATOM 3517 CB LEU 573 40.224 43.91075.317 1.00 2.00 0 ATOM 3518 CG LEU 573 40.807 43.360 76.625 1.00 2.00 0ATOM 3519 CD1 LEU 573 40.667 44.416 77.717 1.00 2.00 0 ATOM 3520 CD2 LEU573 40.079 42.053 77.033 1.00 2.00 0 ATOM 3521 C LEU 573 39.055 43.34473.170 1.00 4.08 0 ATOM 3522 O LEU 573 37.865 42.974 73.260 1.00 2.00 0ATOM 3523 N ARG 574 39.518 44.153 72.203 1.00 13.12 0 ATOM 3525 CA ARG574 38.682 44.665 71.115 1.00 12.30 0 ATOM 3526 CB ARG 574 39.491 45.59870.212 1.00 30.04 0 ATOM 3527 CG ARG 574 39.704 46.994 70.757 1.00 31.000 ATOM 3528 CD ARG 574 40.697 47.811 69.915 1.00 35.07 0 ATOM 3529 NEARG 574 40.294 47.989 68.514 1.00 36.10 0 ATOM 3531 CZ ARG 574 41.01848.625 67.587 1.00 36.14 0 ATOM 3532 NH1 ARG 574 42.198 49.162 67.8901.00 35.84 0 ATOM 3535 NH2 ARG 574 40.565 48.713 66.341 1.00 40.46 0ATOM 3538 C ARG 574 38.265 43.440 70.324 1.00 10.99 0 ATOM 3539 O ARG574 37.092 43.253 70.004 1.00 30.16 0 ATOM 3540 N LEU 575 39.260 42.59170.066 1.00 2.00 0 ATOM 3542 CA LEU 575 39.156 41.339 69.323 1.00 2.00 0ATOM 3543 CB LEU 575 40.471 40.576 69.502 1.00 18.14 0 ATOM 3544 CG LEU575 41.058 39.569 68.514 1.00 18.14 0 ATOM 3545 CD1 LEU 575 40.11238.378 68.370 1.00 18.14 0 ATOM 3546 CD2 LEU 575 41.355 40.249 67.1761.00 18.14 0 ATOM 3547 C LEU 575 37.970 40.494 69.801 1.00 2.00 0 ATOM3548 O LEU 575 37.121 40.081 69.001 1.00 18.14 0 ATOM 3549 N PHE 57637.908 40.247 71.103 1.00 29.25 0 ATOM 3551 CA PHE 576 36.823 39.45971.664 1.00 32.60 0 ATOM 3552 CB PHE 576 37.115 39.098 73.119 1.00 2.000 ATOM 3553 CG PHE 576 38.116 38.001 73.270 1.00 2.00 0 ATOM 3554 CD1PHE 576 39.158 36.111 74.184 1.00 2.00 0 ATOM 3555 CD2 PHE 576 38.02236.855 72.491 1.00 2.00 0 ATOM 3556 CE1 PHE 576 40.097 37.098 74.3241.00 2.00 0 ATOM 3557 CE2 PHE 576 38.960 35.828 72.621 1.00 2.00 0 ATOM3559 CZ PHE 576 40.000 35.953 73.540 1.00 2.00 0 ATOM 3559 C PHE 57635.479 40.164 71.573 1.00 29.95 0 ATOM 3560 O PHE 576 34.433 39.51771.648 1.00 2.00 0 ATOM 3561 N GLU 577 35.504 41.484 71.421 1.00 21.04 0ATOM 3563 CA GLU 577 34.273 42.259 71.307 1.00 23.59 0 ATOM 3564 CB GLU577 34.561 43.731 71.566 1.00 40.21 0 ATOM 3565 CG GLU 577 35.032 44.00172.971 1.00 50.88 0 ATOM 3566 CD GLU 577 35.688 45.347 73.113 1.00 55.670 ATOM 3567 OE1 GLU 577 36.439 45.522 74.099 1.00 60.01 0 ATOM 3568 OE2GLU 577 35.461 46.224 72.245 1.00 63.14 0 ATOM 3569 C GLU 577 33.65442.091 69.928 1.00 25.67 0 ATOM 3570 O GLU 577 32.435 42.156 69.772 1.0044.83 0 ATOM 3571 N TYR 578 34.509 41.901 68.928 1.00 51.92 0 ATOM 3573CA TYR 578 34.052 41.700 67.561 1.00 48.79 0 ATOM 3574 CB TYR 578 35.20841.834 66.564 1.00 35.56 0 ATOM 3575 CG TYR 578 35.703 43.245 66.3561.00 40.25 0 ATOM 3576 CD1 TYR 578 35.598 43.865 65.111 1.00 40.27 0ATOM 3577 CE1 TYR 578 36.062 45.168 64.917 1.00 44.74 0 ATOM 3578 CD2TYR 578 36.281 43.960 67.399 1.00 45.71 0 ATOM 3579 CE2 TYR 578 36.74745.258 67.217 1.00 42.53 0 ATOM 3580 CZ TYR 578 36.636 45.853 65.9791.00 49.98 0 ATOM 3581 OH TYR 578 37.106 47.131 65.812 1.00 54.82 0 ATOM3583 C TYR 578 33.467 40.303 67.458 1.00 49.18 0 ATOM 3584 O TYR 57832.316 40.124 67.081 1.00 35.27 0 ATOM 3585 N GLY 579 34.271 39.30767.797 1.00 2.60 0 ATOM 3587 CA GLY 579 33.787 37.949 67.713 1.00 6.22 0ATOM 3588 C GLY 579 32.737 37.628 68.755 1.00 8.52 0 ATOM 3589 O GLY 57931.555 37.446 68.437 1.00 39.88 0 ATOM 3590 N GLY 580 33.194 37.55770.006 1.00 2.00 0 ATOM 3592 CA GLY 580 32.342 37.236 71.140 1.00 2.00 0ATOM 3593 C GLY 580 33.191 36.491 72.151 1.00 2.00 0 ATOM 3594 O GLY 58033.725 35.415 71.853 1.00 44.94 0 ATOM 3595 N PHE 581 33.307 37.06773.348 1.00 15.06 0 ATOM 3597 CA PHE 581 34.116 36.500 74.432 1.00 12.630 ATOM 3598 CB PHE 581 33.857 37.263 75.744 1.00 2.00 0 ATOM 3599 CG PHE581 34.679 38.542 75.885 1.00 2.00 0 ATOM 3600 CD1 PHE 581 34.136 39.78375.557 1.00 2.00 0 ATOM 3601 CD2 PHE 581 35.999 38.495 76.346 1.00 2.000 ATOM 3602 CE1 PHE 581 34.894 40.941 75.687 1.00 2.00 0 ATOM 3603 CE2PHE 581 36.757 39.659 76.475 1.00 2.00 0 ATOM 3604 CZ PHE 581 36.20440.875 76.146 1.00 2.00 0 ATOM 3605 C PHE 581 33.913 34.999 74.611 1.0012.63 0 ATOM 3606 O PHE 581 32.782 34.521 74.647 1.00 2.00 0 ATOM 3607 NPRO 582 35.021 34.243 74.737 1.00 21.81 0 ATOM 3608 CD PRO 582 36.36034.852 74.820 1.00 85.20 0 ATOM 3609 CA PRO 582 35.141 32.795 74.9101.00 24.62 0 ATOM 3610 CP PRO 582 36.398 32.675 75.737 1.00 86.40 0 ATOM3611 CG PRO 582 37.266 33.636 75.008 1.00 87.84 0 ATOM 3612 C PRO 58233.966 31.958 75.425 1.00 28.53 0 ATOM 3613 O PRO 582 33.816 30.79675.012 1.00 0.58 0 ATOM 3614 N PRO 583 33.160 32.466 76.374 1.00 13.00 0ATOM 3615 CD PRO 583 33.090 33.632 77.269 1.00 51.24 0 ATOM 3616 CA PRO583 32.099 31.500 76.681 1.00 13.55 0 ATOM 3617 CB PRO 583 31.281 32.21577.759 1.00 53.59 0 ATOM 3618 CG PRO 583 31.616 33.699 77.550 1.00 50.460 ATOM 3619 C PRO 583 31.296 31.283 75.389 1.00 14.03 0 ATOM 3620 O PRO583 30.950 30.152 75.043 1.00 48.29 0 ATOM 3621 N GLU 584 31.089 32.38374.663 1.00 35.81 0 ATOM 3623 CA GLU 584 30.333 32.439 73.412 1.00 39.440 ATOM 3624 CB GLU 584 30.122 33.913 73.043 1.00 78.69 0 ATOM 3625 CGGLU 584 28.955 34.213 72.117 1.00 90.10 0 ATOM 3626 CD GLU 584 28.66235.712 72.022 1.00 95.41 0 ATOM 3627 OE1 GLU 584 28.313 36.190 70.9181.00 90.41 0 ATOM 3628 OE2 GLU 584 28.782 36.418 73.053 1.00 95.61 0ATOM 3629 C GLU 584 30.975 31.676 72.244 1.00 36.17 0 ATOM 3630 O GLU584 30.780 30.466 72.106 1.00 78.39 0 ATOM 3631 N SER 585 31.737 32.38171.406 1.00 12.28 0 ATOM 3633 CA SER 585 32.394 31.779 70.245 1.00 6.770 ATOM 3634 CD SER 585 32.720 32.857 69.209 1.00 31.98 0 ATOM 3635 OGSER 585 31.558 33.556 68.809 1.00 27.38 0 ATOM 3637 C SER 585 33.66930.995 70.539 1.00 9.84 0 ATOM 3638 O SER 585 34.382 31.258 71.502 1.0035.35 0 ATOM 3639 N ASN 586 33.947 30.037 69.664 1.00 2.00 0 ATOM 3641CA ASN 586 35.136 29.197 69.749 1.00 2.00 0 ATOM 3642 CB ASN 586 34.90927.854 69.066 1.00 2.87 0 ATOM 3643 CG ASN 586 33.728 27.107 69.625 1.0010.37 0 ATOM 3644 OD1 ASN 586 33.497 25.960 69.265 1.00 11.13 0 ATOM3645 ND2 ASN 586 32.967 27.745 70.506 1.00 4.44 0 ATOM 3648 C ASN 58636.251 29.937 69.029 1.00 2.00 0 ATOM 3649 O ASN 586 36.017 30.57468.007 1.00 2.87 0 ATOM 3650 N TYR 587 37.458 29.858 69.564 1.00 13.99 0ATOM 3652 CA TYR 587 38.571 30.554 68.966 1.00 13.99 0 ATOM 3653 CB TYR587 39.049 31.706 69.876 1.00 2.00 0 ATOM 3654 CG TYR 587 38.177 32.94669.873 1.00 2.00 0 ATOM 3655 CD1 TYR 587 36.998 33.004 70.638 1.00 2.000 ATOM 3656 CE1 TYR 587 36.177 34.138 70.624 1.00 2.00 0 ATOM 3657 CD2TYR 587 38.519 34.061 69.088 1.00 2.00 0 ATOM 3658 CE2 TYR 587 37.70135.205 69.065 1.00 2.00 0 ATOM 3659 CZ TYR 587 36.532 35.231 69.840 1.002.00 0 ATOM 3660 OH TYR 587 35.743 36.352 69.838 1.00 2.00 0 ATOM 3662 CTYR 587 39.721 29.608 68.745 1.00 13.99 0 ATOM 3663 O TYR 587 39.81228.562 69.392 1.00 2.00 0 ATOM 3664 N LEU 586 40.584 29.987 67.809 1.002.00 0 ATOM 3666 CA LEU 588 41.803 29.261 67.493 1.00 2.00 0 ATOM 3667CB LEU 588 41.681 29.482 66.195 1.00 10.69 0 ATOM 3668 CG LEU 588 43.03227.907 65.781 1.00 10.69 0 ATOM 3669 CD1 LEU 588 43.568 27.073 66.9121.00 10.69 0 ATOM 3670 CD2 LEU 588 42.912 27.079 64.533 1.00 10.69 0ATOM 3671 C LEU 588 42.809 30.375 67.301 1.00 2.00 0 ATOM 3672 O LEU 58842.547 31.301 66.532 1.00 10.69 0 ATOM 3673 N PHE 589 43.924 30.33366.026 1.00 2.00 0 ATOM 3675 CA PHE 589 44.938 31.363 67.865 1.00 2.00 0ATOM 3676 CB PHE 589 45.289 32.013 69.200 1.00 2.00 0 ATOM 3677 CG PHE589 44.279 33.026 69.652 1.00 2.00 0 ATOM 3678 CD1 PHE 589 43.136 32.63370.326 1.00 2.00 0 ATOM 3679 CD2 PHE 589 44.471 34.376 69.391 1.00 2.000 ATOM 3680 CE1 PHE 589 42.203 33.567 70.732 1.00 2.00 0 ATOM 3681 CE2PHE 589 43.542 35.321 69.793 1.00 2.00 0 ATOM 3682 CZ PHE 589 42.40834.918 70.463 1.00 2.00 0 ATOM 3683 C PHE 589 46.140 30.717 67.219 1.002.00 0 ATOM 3684 O PHE 589 46.532 29.610 67.602 1.00 2.00 0 ATOM 3685 NLEU 590 46.715 31.411 66.235 1.00 2.00 0 ATOM 3687 CA LEU 590 47.84530.892 65.459 1.00 2.00 0 ATOM 3688 CB LEU 590 47.650 31.303 63.988 1.009.73 0 ATOM 3689 CG LEU 590 46.273 31.022 63.356 1.00 9.73 0 ATOM 3690CD1 LEU 590 46.162 31.702 62.014 1.00 9.73 0 ATOM 3691 CD2 LEU 59046.056 29.529 63.211 1.00 9.73 0 ATOM 3692 C LEU 590 49.277 31.22165.947 1.00 2.00 0 ATOM 3693 O LEU 590 50.263 30.856 65.296 1.00 9.73 0ATOM 3694 N GLY 591 49.389 31.929 67.071 1.00 2.00 0 ATOM 3696 CA GLY591 50.703 32.223 67.626 1.00 2.00 0 ATOM 3697 C GLY S91 51.132 33.64867.923 1.00 2.00 0 ATOM 3698 O GLY 591 50.369 34.620 67.769 1.00 2.00 0ATOM 3699 N ASP 592 52.387 33.741 68.358 1.00 2.00 0 ATOM 3701 CA ASP592 53.033 35.000 68.707 1.00 2.00 0 ATOM 3702 CB ASP 592 53.424 35.74667.448 1.00 6.15 a ATOM 3703 CG ASP 592 54.521 35.051 66.686 1.00 16.840 ATOM 3704 OD1 ASP 592 54.955 35.606 65.657 1.00 11.71 0 ATOM 3705 OD2ASP S92 54.943 33.949 67.114 1.00 18.80 0 ATOM 3706 C ASP 592 52.19435.887 69.598 1.00 2.00 0 ATOM 3707 O ASP 592 51.813 36.995 69.211 1.006.68 0 ATOM 3708 N TYR 593 51.927 35.370 70.800 1.00 12.61 0 ATOM 3710CA TYR 593 51.127 36.036 71.822 1.00 12.61 0 ATOM 3711 CB TYR 593 50.60235.017 72.827 1.00 2.00 0 ATOM 3712 CG TYR 593 49.994 33.790 72.200 1.002.00 0 ATOM 3713 CD1 TYR 593 50.558 32.532 72.382 1.00 2.00 0 ATOM 3714CE1 TYR 593 49.995 31.401 71.793 1.00 2.00 0 ATOM 3715 CD2 TYR 59348.852 33.886 71.413 1.00 2.00 0 ATOM 3716 CE2 TYR 593 48.284 32.77270.822 1.00 2.00 0 ATOM 3717 CZ TYR 593 48.856 31.540 71.013 1.00 2.00 0ATOM 3718 OH TYR 593 48.267 30.459 70.412 1.00 2.00 0 ATOM 3720 C TYR593 51.995 37.019 72.560 1.00 12.61 0 ATOM 3721 O TYR 593 51.526 38.07972.981 1.00 2.00 0 ATOM 3722 N VAL 594 53.270 36.660 72.692 1.00 2.00 0ATOM 3724 CA VAL 594 54.238 37.463 73.420 1.00 2.00 0 ATOM 3725 CB VAL594 54.913 36.599 74.501 1.00 30.06 0 ATOM 3726 CG1 VAL 594 53.85535.785 75.241 1.00 30.06 0 ATOM 3727 CG2 VAL 594 55.950 35.684 73.8751.00 30.06 0 ATOM 3728 C VAL 594 55.309 38.154 72.563 1.00 2.00 0 ATOM3729 O VAL 594 55.408 37.919 71.356 1.00 30.06 0 ATOM 3730 N ASP 59556.112 38.991 73.218 1.00 2.00 0 ATOM 3732 CA ASP 595 57.184 39.77672.601 1.00 2.00 0 ATOM 3733 CB ASP 595 58.073 38.903 71.705 1.00 33.250 ATOM 3734 CG ASP 595 58.911 37.890 72.502 1.00 42.03 0 ATOM 3735 OD1ASP 595 59.240 38.147 73.679 1.00 40.72 0 ATOM 3736 OD2 ASP 595 59.25636.831 71.941 1.00 46.45 0 ATOM 3737 C ASP 595 56.547 40.920 71.818 1.002.00 0 ATOM 3738 O ASP 595 55.335 40.901 71.575 1.00 29.29 0 ATOM 3739 NARG 596 57.342 41.927 71.460 1.00 2.00 0 ATOM 3741 CA ARG 596 56.84343.096 70.716 1.00 2.00 0 ATOM 3742 CB ARG 596 56.146 42.658 69.416 1.007.10 0 ATOM 3743 CG ARG 596 57.043 41.863 68.457 1.00 13.06 0 ATOM 3744CD ARG 596 58.035 42.755 67.729 1.00 17.94 0 ATOM 3745 NE ARG 596 59.29342.081 67.407 1.00 25.56 0 ATOM 3747 CZ ARG 596 59.402 40.954 66.7091.00 35.06 0 ATOM 3748 NH1 ARG 596 58.325 40.338 66.236 1.00 31.67 0ATOM 3751 NH2 ARG 596 60.604 40.438 66.486 1.00 31.81 0 ATOM 3754 C ARG596 55.898 43.996 71.547 1.00 2.00 0 ATOM 3755 O ARG 596 56.269 45.11271.922 1.00 7.10 0 ATOM 3756 N GLY 597 54.692 43.521 71.847 1.00 32.20 0ATOM 3758 CA GLY 597 53.763 44.323 72.632 1.00 30.86 0 ATOM 3759 C GLY597 54.199 44.603 74.062 1.00 35.68 0 ATOM 3760 O GLY 597 55.120 43.97674.570 1.00 2.00 0 ATOM 3761 N LYS 598 53.520 45.539 74.720 1.00 20.71 0ATOM 3763 CA LYS 598 53.845 45.911 76.097 1.00 17.90 0 ATOM 3764 CB LYS598 53.686 47.427 76.271 1.00 54.79 0 ATOM 3765 CG LYS 598 54.813 48.25975.655 1.00 53.06 0 ATOM 3766 CD LYS 598 56.159 48.031 76.359 1.00 56.270 ATOM 3767 CE LYS 598 56.120 48.441 77.838 1.00 53.59 0 ATOM 3768 NZLYS 598 57.407 48.186 78.548 1.00 56.50 0 ATOM 3772 C LYS 598 53.06445.179 77.210 1.00 18.19 0 ATOM 3773 O LYS 598 53.384 45.314 78.395 1.0055.50 0 ATOM 3774 N GLN 599 52.053 44.400 76.832 1.00 2.00 0 ATOM 3776CA GLN 599 51.244 43.666 77.795 1.00 2.00 0 ATOM 3777 CB GLN 599 49.82044.244 77.839 1.00 24.44 0 ATOM 3778 CG GLN 599 49.780 45.701 78.3041.00 31.08 0 ATOM 3779 CD GLN 599 48.398 46.169 78.728 1.00 27.33 0 ATOM3780 OE1 GLN 599 47.537 46.449 77.895 1.00 26.31 0 ATOM 3781 NE2 GLN 59948.188 46.277 80.029 1.00 27.77 0 ATOM 3784 C GLN 599 51.215 42.16377.514 1.00 2.00 0 ATOM 3785 O GLN 599 50.183 41.515 77.626 1.00 24.92 0ATOM 3786 N SER 600 52.371 41.602 77.198 1.00 2.00 0 ATOM 3788 CA SER600 52.445 40.180 76.921 1.00 2.00 0 ATOM 3789 CD SER 600 53.975 39.78176.547 1.00 2.00 0 ATOM 3790 OG SER 600 54.381 40.577 75.480 1.00 2.00 0ATOM 3792 C SER 600 51.968 39.304 78.078 1.00 2.00 0 ATOM 3793 O SER 60051.589 38.153 77.856 1.00 2.00 0 ATOM 3794 N LEU 601 51.990 39.83079.306 1.00 2.00 0 ATOM 3796 CA LEU 601 51.566 39.055 80.486 1.00 2.00 0ATOM 3797 CB LEU 601 52.120 39.628 81.801 1.00 2.00 0 ATOM 3798 CG LEU601 53.573 39.753 62.260 1.00 2.00 0 ATOM 3791 CD1 LEU 601 54.290 38.40582.228 1.00 2.00 0 ATOM 3800 CD2 LEU 601 54.244 40.799 81.399 1.00 2.000 ATOM 3801 C LEU 601 50.058 39.002 80.635 1.00 2.00 0 ATOM 3802 O LEU601 49.498 37.949 80.921 1.00 4.13 0 ATOM 3803 N GLU 602 49.412 40.15480.490 1.00 2.00 0 ATOM 3805 CA GLU 602 47.969 40.227 80.608 1.00 2.00 0ATOM 3806 CB GLU 602 47.486 41.676 80.495 1.00 5.18 0 ATOM 3807 CG GLU602 47.752 42.552 81.739 1.00 5.18 0 ATOM 3808 CD GLU 602 49.152 43.18081.773 1.00 5.18 0 ATOM 3809 OE1 GLU 602 50.146 42.439 81.854 1.00 5.180 ATOM 3810 OE2 GLU 602 49.266 44.424 81.732 1.00 5.18 0 ATOM 3811 C GLU602 47.413 39.391 79.479 1.00 2.00 0 ATOM 3812 O GLU 602 46.452 38.66179.661 1.00 5.18 0 ATOM 3813 N THR 603 48.072 39.477 78.323 1.00 56.10 0ATOM 3815 CA THR 603 47.705 38.748 77.110 1.00 56.10 0 ATOM 3816 CB THR603 48.585 39.206 75.941 1.00 8.67 0 ATOM 3817 OG1 THR 603 48.197 40.52775.557 1.00 8.67 0 ATOM 3819 CG2 THR 603 48.437 38.272 74.748 1.00 8.670 ATOM 3820 C THR 603 47.794 37.228 77.228 1.00 56.10 0 ATOM 3821 O THR603 46.804 36.522 77.060 1.00 8.67 0 ATOM 3822 N ILE 604 48.980 36.71377.505 1.00 2.00 0 ATOM 3824 CA ILE 604 49.137 35.273 77.630 1.00 2.00 0ATOM 3825 CB ILE 604 50.643 34.900 77.738 1.00 2.00 0 ATOM 3826 CG2 ILE604 51.244 35.475 79.009 1.00 2.00 0 ATOM 3827 CG1 ILE 604 50.812 33.38277.627 1.00 2.00 0 ATOM 3828 CD1 ILE 604 50.177 32.766 76.369 1.00 2.000 ATOM 3829 C ILE 604 48.314 34.710 78.808 1.00 2.00 0 ATOM 3830 O ILE604 47.886 33.565 78.779 1.00 2.00 0 ATOM 3831 N CYS 605 48.057 35.52679.821 1.00 17.68 0 ATOM 3833 CA CYS 605 47.283 35.072 80.962 1.00 16.480 ATOM 3834 CB CYS 605 47.434 36.026 82.138 1.00 12.63 0 ATOM 3835 SGCYS 605 48.994 35.843 82.980 1.00 18.12 0 ATOM 3836 C CYS 605 45.82434.923 80.630 1.00 10.22 0 ATOM 3837 O CYS 605 45.185 33.963 81.068 1.008.59 0 ATOM 3838 N LEU 606 45.280 35.856 79.861 1.00 2.00 0 ATOM 3840 CALEU 606 43.874 35.771 79.504 1.00 2.00 0 ATOM 3841 CB LEU 606 43.39637.087 78.881 1.00 2.00 0 ATOM 3842 CG LEU 606 41.886 37.223 78.636 1.002.00 0 ATOM 3843 CD1 LEU 606 41.081 36.716 79.867 1.00 2.00 0 ATOM 3844CD2 LEU 606 41.551 38.688 78.326 1.00 2.00 0 ATOM 3845 C LEU 606 43.58934.599 78.559 1.00 2.00 0 ATOM 3846 O LEU 606 42.503 34.012 78.612 1.002.00 0 ATOM 3847 N LEU 607 44.562 34.246 77.713 1.00 9.59 0 ATOM 3849 CALEU 607 44.392 33.137 76.772 1.00 9.59 0 ATOM 3850 CB LEU 607 45.39433.246 75.604 1.00 9.66 0 ATOM 3851 CG LEU 607 45.302 34.531 74.755 1.009.66 0 ATOM 3852 CD1 LEU 607 46.376 34.564 73.709 1.00 9.66 0 ATOM 3853CD2 LEU 607 43.951 34.634 74.114 1.00 9.66 0 ATOM 3854 C LEU 607 44.50931.781 77.476 1.00 9.59 0 ATOM 3855 O LEU 607 43.772 30.841 77.148 1.009.66 0 ATOM 3856 N LEU 608 45.412 31.688 78.452 1.00 67.56 0 ATOM 3858CA LEU 608 45.599 30.460 79.230 1.00 67.56 0 ATOM 3859 CB LEU 608 46.87230.529 80.054 1.00 2.00 0 ATOM 3860 CG LEU 608 48.168 30.385 79.273 1.002.00 0 ATOM 3861 CD1 LEU 608 49.357 30.366 80.234 1.00 2.00 0 ATOM 3862CD2 LEU 608 48.109 29.107 78.462 1.00 2.00 0 ATOM 3863 C LEU 608 44.42730.217 80.170 1.00 67.56 0 ATOM 3864 O LEU 608 44.097 29.059 50.477 1.002.00 0 ATOM 3865 N ALA 609 43.833 31.313 60.653 1.00 2.00 0 ATOM 3867 CAALA 609 42.667 31.250 81.531 1.00 2.00 0 ATOM 3868 CB ALA 609 42.32232.637 82.044 1.00 2.00 0 ATOM 3869 C ALA 609 41.496 30.677 80.734 1.002.00 0 ATOM 3870 O ALA 609 40.833 29.747 81.162 1.00 2.00 0 ATOM 3871 NTYR 610 41.263 31.231 79.552 1.00 2.00 0 ATOM 3873 CA TYR 610 40.18630.771 78.686 1.00 2.00 0 ATOM 3874 CB TYR 610 40.046 31.717 77.504 1.002.00 0 ATOM 3875 CG TYR 610 39.274 32.974 77.820 1.00 2.00 0 ATOM 3876CD1 TYR 610 39.685 34.202 77.315 1.00 2.00 0 ATOM 3877 CE1 TYR 61038.952 35.359 77.558 1.00 2.00 0 ATOM 3878 CD2 TYR 610 38.108 32.93378.589 1.00 2.00 0 ATOM 3879 CE2 TYR 610 37.367 34.091 78.841 1.00 2.000 ATOM 3880 CZ TYR 610 37.797 35.298 78.319 1.00 2.00 0 ATOM 3881 OH TYR610 37.086 36.452 78.533 1.00 2.00 0 ATOM 3883 C TYR 610 40.431 29.34278.205 1.00 2.00 0 ATOM 3884 O TYR 610 39.481 28.575 78.006 1.00 2.00 0ATOM 3885 N LYS 611 41.703 26.991 78.017 1.00 2.00 0 ATOM 3887 CA LYS611 42.063 27.648 77.600 1.00 2.00 0 ATOM 3888 CB LYS 611 43.551 27.53277.308 1.00 3.53 0 ATOM 3889 CG LYS 611 43.926 26.136 76.803 1.00 3.53 0ATOM 3890 CD LYS 611 43.240 25.837 75.467 1.00 3.53 0 ATOM 3891 CE LYS611 43.476 24.412 74.980 1.00 3.53 0 ATOM 3892 NZ LYS 611 42.391 23.50275.421 1.00 3.53 0 ATOM 3896 C LYS 611 41.717 26.666 76.700 1.00 2.00 0ATOM 3897 O LYS 611 41.142 25.616 78.431 1.00 3.53 0 ATOM 3898 N ILE 61242.084 26.990 79.938 1.00 26.02 0 ATOM 3900 CA ILE 612 41.780 26.11581.069 1.00 26.02 0 ATOM 3901 CB ILE 612 42.543 26.535 82.336 1.00 2.000 ATOM 3902 CG2 ILE 612 42.232 25.576 83.475 1.00 2.00 0 ATOM 3903 CG1ILE 612 44.041 26.511 82.063 1.00 2.00 0 ATOM 3904 CD1 ILE 612 44.86027.144 83.135 1.00 2.00 0 ATOM 3905 C ILE 612 40.284 26.149 81.365 1.0026.02 0 ATOM 3906 O ILE 612 39.698 25.131 81.739 1.00 2.00 0 ATOM 3907 NLYS 613 39.683 27.326 81.181 1.00 2.00 0 ATOM 3909 CA LYS 613 38.25727.556 81.414 1.00 2.00 0 ATOM 3910 CD LYS 613 37.966 29.056 81.402 1.0010.53 0 ATOM 3911 CG LYS 613 36.528 29.437 81.650 1.00 10.53 0 ATOM 3912CD LYS 613 36.070 29.072 83.035 1.00 10.53 0 ATOM 3913 CE LYS 613 34.71729.693 83.364 1.00 10.53 0 ATOM 3914 NZ LYS 613 33.611 29.182 82.5231.00 10.53 0 ATOM 3918 C LYS 613 37.379 26.847 80.384 1.00 2.00 0 ATOM3919 O LYS 613 36.335 26.293 80.739 1.00 10.53 0 ATOM 3920 N TYR 61437.819 26.842 79.121 1.00 24.09 0 ATOM 3922 CA TYR 614 37.079 26.21478.016 1.00 27.84 0 ATOM 3923 CB TYR 614 36.473 27.301 77.125 1.00 2.000 ATOM 3924 CG TYR 614 35.679 26.363 77.855 1.00 2.00 0 ATOM 3925 CD1TYR 614 36.123 29.689 77.892 1.00 2.00 0 ATOM 3926 CE1 TYR 614 35.40930.671 78.567 1.00 2.00 0 ATOM 3927 CD2 TYR 614 34.490 28.048 78.5161.00 2.00 0 ATOM 3928 CE2 TYR 614 33.766 29.020 79.199 1.00 2.00 0 ATOM3929 CZ TYR 614 34.232 30.330 79.227 1.00 2.00 0 ATOM 3930 OH TYR 61433.560 31.293 79.960 1.00 2.00 0 ATOM 3932 C TYR 614 37.976 25.31777.146 1.00 25.93 0 ATOM 3933 O TYR 614 38.012 25.474 75.935 1.00 2.00 0ATOM 3934 N PRO 615 38.641 24.310 77.733 1.00 21.49 0 ATOM 3935 CD PRO615 38.494 23.854 79.123 1.00 11.83 0 ATOM 3936 CA PRO 615 39.541 23.40676.995 1.00 22.47 0 ATOM 3937 CB PRO 615 39.950 22.384 78.055 1.00 11.830 ATOM 3938 CG PRO 615 38.773 22.382 78.995 1.00 11.83 0 ATOM 3939 C PRO615 39.078 22.714 75.720 1.00 20.23 0 ATOM 3940 O PRO 615 39.873 22.02675.074 1.00 11.83 0 ATOM 3941 N GLU 616 37.813 22.879 75.350 1.00 15.620 ATOM 3943 CA GLU 616 37.296 22.213 74.159 1.00 16.61 0 ATOM 3944 CBGLU 616 36.240 21.179 74.566 1.00 26.75 0 ATOM 3945 CG GLU 616 36.64420.253 75.695 1.00 30.68 0 ATOM 3946 CD GLU 616 37.872 19.416 75.3821.00 40.71 0 ATOM 3947 OE1 GLU 616 38.747 19.294 76.273 1.00 39.65 0ATOM 3948 OE2 GLU 616 37.960 18.874 74.256 1.00 39.29 0 ATOM 3949 C GLU616 36.681 23.175 73.140 1.00 13.01 0 ATOM 3950 O GLU 616 36.276 22.76572.046 1.00 20.06 0 ATOM 3951 N ASN 617 36.620 24.453 73.499 1.00 2.00 0ATOM 3953 CA ASN 617 36.027 25.467 72.636 1.00 2.00 0 ATOM 3954 CB ASN617 34.678 25.896 73.218 1.00 42.63 0 ATOM 3955 CG ASN 617 33.743 24.72073.456 1.00 47.29 0 ATOM 3956 OD1 ASN 617 33.800 24.064 74.500 1.0050.83 0 ATOM 3957 ND2 ASN 617 32.883 24.443 72.485 1.00 53.65 0 ATOM3960 C ASN 617 36.941 26.686 72.476 1.00 2.00 0 ATOM 3961 O ASN 61736.500 27.747 72.032 1.00 45.01 0 ATOM 3962 N PHE 618 38.215 26.51772.812 1.00 14.67 0 ATOM 3964 CA PHE 618 39.192 27.596 72.736 1.00 14.670 ATOM 3965 CB PHE 618 39.146 28.387 74.044 1.00 2.00 0 ATOM 3966 CG PHE618 40.042 29.585 74.073 1.00 2.00 0 ATOM 3967 CD1 PHE 618 39.501 30.87174.058 1.00 2.00 0 ATOM 3968 CD2 PHE 618 41.422 29.441 74.147 1.00 2.000 ATOM 3969 CE1 PHE 618 40.322 32.007 74.119 1.00 2.00 0 ATOM 3970 CE2PHE 618 42.254 30.562 74.209 1.00 2.00 0 ATOM 3971 CZ PHE 618 41.69631.854 74.195 1.00 2.00 0 ATOM 3972 C PHE 618 40.540 26.907 72.570 1.0014.67 0 ATOM 3973 O PHE 618 40.893 26.066 73.394 1.00 2.00 0 ATOM 3974 NPHE 619 41.285 27.239 71.514 1.00 2.00 0 ATOM 3976 CA PHE 619 42.58326.599 71.271 1.00 2.00 0 ATOM 3977 CB PHE 619 42.468 25.622 70.111 1.002.00 0 ATOM 3978 CG PHE 619 41.404 24.595 70.311 1.00 2.00 0 ATOM 3979CD1 PHE 619 40.063 24.936 70.159 1.00 2.00 0 ATOM 3980 CD2 PHE 61941.738 23.291 70.678 1.00 2.00 0 ATOM 3981 CE1 PHE 619 39.068 23.99770.370 1.00 2.00 0 ATOM 3982 CE2 PHE 619 40.750 22.337 70.892 1.00 2.000 ATOM 3983 CZ PHE 619 39.408 22.689 70.738 1.00 2.00 0 ATOM 3984 C PHE619 43.719 27.566 71.005 1.00 2.00 0 ATOM 3985 O PHE 619 43.501 26.69870.564 1.00 2.00 0 ATOM 3986 N LEU 620 44.936 27.115 71.282 1.00 2.00 0ATOM 3988 CA LEU 620 46.118 27.933 71.073 1.00 2.00 0 ATOM 3989 CB LEU620 46.647 28.507 72.393 1.00 2.00 0 ATOM 3990 CG LEU 620 45.826 29.38973.321 1.00 2.00 0 ATOM 3991 CD1 LEU 620 46.741 29.868 74.437 1.00 2.000 ATOM 3992 CD2 LEU 620 45.247 30.563 72.565 1.00 2.00 0 ATOM 3993 C LEU620 47.225 27.093 70.438 1.00 2.00 0 ATOM 3994 O LEU 620 47.548 25.98770.910 1.00 2.00 0 ATOM 3995 N LEU 621 47.805 27.634 69.376 1.00 2.00 0ATOM 3997 CA LEU 621 48.880 26.971 68.669 1.00 2.00 0 ATOM 3998 CB LEU621 48.577 26.929 67.162 1.00 2.00 0 ATOM 3999 CG LEU 621 47.256 26.28066.739 1.00 2.00 0 ATOM 4000 CD1 LEU 621 47.021 26.511 65.253 1.00 2.000 ATOM 4001 CD2 LEU 621 47.281 24.813 67.062 1.00 2.00 0 ATOM 4002 C LEU621 50.162 27.756 68.943 1.00 2.00 0 ATOM 4003 O LEU 621 50.121 28.93769.285 1.00 2.00 0 ATOM 4004 N ARG 622 51.296 27.092 68.774 1.00 2.00 0ATOM 4006 CA ARG 622 52.593 27.690 69.013 1.00 2.00 0 ATOM 4007 CB ARG622 53.620 26.578 69.231 1.00 5.18 0 ATOM 4008 CG ARG 622 54.967 27.05269.716 1.00 5.18 0 ATOM 4009 CD ARG 622 55.802 25.891 70.196 1.00 5.18 0ATOM 4010 NE ARG 622 56.969 26.357 70.941 1.00 5.18 0 ATOM 4012 CZ ARG622 57.830 25.553 71.557 1.00 5.18 0 ATOM 4013 NH1 ARG 622 57.659 24.24071.525 1.00 6.18 0 ATOM 4016 NH2 ARG 622 58.870 26.056 72.199 1.00 5.180 ATOM 4019 C ARG 622 53.070 28.615 67.894 1.00 2.00 0 ATOM 4020 O ARG622 53.032 28.270 66.702 1.00 16.40 0 ATOM 4021 N GLY 623 53.508 29.80568.286 1.00 19.04 0 ATOM 4023 CA GLY 623 54.043 30.749 67.328 1.00 20.780 ATOM 4024 C GLY 623 55.551 30.573 67.390 1.00 18.09 0 ATOM 4025 O GLY623 56.069 29.805 68.212 1.00 2.00 0 ATOM 4026 N ASN 624 56.281 31.26666.530 1.00 32.06 0 ATOM 4028 CA ASN 624 57.728 31.144 66.566 1.00 32.950 ATOM 4029 CB ASN 624 58.345 31.543 65.220 1.00 12.63 0 ATOM 4030 CGASN 624 58.120 33.002 64.857 1.00 8.84 0 ATOM 4031 OD1 ASN 624 57.38133.754 65.538 1.00 9.47 0 ATOM 4032 ND2 ASN 624 58.721 33.413 63.7571.00 8.50 0 ATOM 4035 C ASN 624 58.260 31.988 67.721 1.00 32.42 0 ATOM4036 O ASN 624 59.442 31.926 68.066 1.00 7.81 0 ATOM 4037 N HIS 62557.359 32.757 68.335 1.00 7.71 0 ATOM 4039 CA HIS 625 57.687 33.61469.477 1.00 8.83 0 ATOM 4040 CB HIS 625 57.030 34.987 69.320 1.00 2.00 0ATOM 4041 CG HIS 625 57.909 35.998 68.656 1.00 2.00 0 ATOM 4042 CD2 HIS625 59.042 35.854 67.933 1.00 2.00 0 ATOM 4043 ND1 HIS 625 57.668 37.35168.721 1.00 2.00 0 ATOM 4045 CE1 HIS 625 58.618 37.999 68.069 1.00 2.000 ATOM 4046 NE2 HIS 625 59.464 37.113 67.582 1.00 2.00 0 ATOM 4048 C HIS625 57.278 32.960 70.807 1.00 8.94 0 ATOM 4049 O HIS 625 57.188 33.64471.825 1.00 2.00 0 ATOM 4050 N GLU 626 56.984 31.691 70.767 1.00 2.00 0ATOM 4052 CA GLU 626 56.629 30.914 71.945 1.00 2.00 0 ATOM 4053 CB GLU626 55.404 30.026 71.666 1.00 23.06 0 ATOM 4054 CG GLU 626 54.051 30.62772.039 1.00 18.70 0 ATOM 4055 CD GLU 626 53.812 31.997 71.445 1.00 17.000 ATOM 4056 OE1 GLU 626 53.516 32.922 72.217 1.00 20.31 0 ATOM 4057 OE2GLU 626 53.911 32.165 70.214 1.00 23.26 0 ATOM 4058 C GLU 626 57.88830.068 72.029 1.00 2.00 0 ATOM 4059 O GLU 626 57.851 28.841 71.921 1.0023.50 0 ATOM 4060 N CYS 627 59.015 30.748 72.184 1.00 2.00 0 ATOM 4062CA CYS 627 60.290 30.067 72.218 1.00 2.00 0 ATOM 4063 CB CYS 627 60.83229.957 70.789 1.00 7.85 0 ATOM 4064 SG CYS 627 62.235 29.843 70.575 1.0019.90 0 ATOM 4065 C CYS 627 61.260 30.842 73.105 1.00 2.00 0 ATOM 4066 OCYS 627 61.355 32.066 73.008 1.00 8.12 0 ATOM 4067 N ALA 628 61.97130.116 73.970 1.00 19.49 0 ATOM 4069 CA ALA 628 62.934 30.702 74.9001.00 19.49 0 ATOM 4070 CB ALA 628 63.704 29.601 75.567 1.00 2.00 0 ATOM4071 C ALA 628 63.890 31.662 74.206 1.00 19.49 0 ATOM 4072 O ALA 62863.921 32.859 74.484 1.00 2.00 0 ATOM 4073 N SER 629 64.660 31.10173.291 1.00 10.83 0 ATOM 4075 CA SER 629 65.637 31.813 72.478 1.00 14.600 ATOM 4076 CB SER 629 66.057 30.877 71.355 1.00 15.99 0 ATOM 4077 OGSER 629 65.846 29.522 71.762 1.00 20.43 0 ATOM 4079 C SER 629 65.11933.131 71.899 1.00 19.16 0 ATOM 4080 O SER 629 65.869 34.085 71.759 1.0013.43 0 ATOM 4081 N ILE 630 63.834 33.167 71.566 1.00 2.00 0 ATOM 4083CA ILE 630 63.198 34.354 71.022 1.00 2.00 0 ATOM 4084 CB ILE 630 62.07633.973 70.017 1.00 2.00 0 ATOM 4085 CG2 ILE 630 61.675 35.174 69.2021.00 2.00 0 ATOM 4086 CG1 ILE 630 62.604 32.998 68.980 1.00 2.00 0 ATOM4087 CD1 ILE 630 63.664 33.598 68.080 1.00 2.00 0 ATOM 4088 C ILE 63062.636 35.281 72.131 1.00 2.00 0 ATOM 4089 O ILE 630 62.866 36.49272.068 1.00 2.00 0 ATOM 4090 N ASN 631 61.932 34.729 73.137 1.00 10.29 0ATOM 4092 CA ASN 631 61.350 35.504 74.272 1.00 9.61 0 ATOM 4093 CB ASN631 60.759 34.597 75.349 1.00 8.67 0 ATOM 4094 CG ASN 631 59.555 33.84874.891 1.00 17.37 0 ATOM 4095 OD1 ASN 631 59.189 33.896 73.722 1.0021.10 0 ATOM 4096 ND2 ASN 631 58.926 33.124 75.815 1.00 15.25 0 ATOM4099 C ASN 631 62.367 36.370 75.003 1.00 6.75 0 ATOM 4100 O ASN 63162.059 37.486 75.438 1.00 6.80 0 ATOM 4101 N ARG 632 63.555 35.81075.189 1.00 2.00 0 ATOM 4103 CA ARG 632 64.635 36.492 75.867 1.00 2.00 0ATOM 4104 CB ARG 632 65.873 35.595 75.909 1.00 6.06 0 ATOM 4105 CG ARG632 66.361 35.244 77.316 1.00 8.20 0 ATOM 4106 CD ARG 632 67.436 36.20277.839 1.00 8.31 0 ATOM 4107 NE ARG 632 67.003 37.598 77.918 1.00 11.750 ATOM 4109 CZ ARG 632 67.837 38.635 77.907 1.00 13.36 0 ATOM 4110 NH1ARG 632 69.144 38.423 77.811 1.00 13.50 0 ATOM 4113 NH2 ARG 632 67.37239.883 78.001 1.00 18.51 0 ATOM 4116 C ARG 632 64.962 37.781 75.148 1.002.00 0 ATOM 4117 O ARG 632 64.930 38.865 75.745 1.00 12.49 0 ATOM 4118 NILE 633 65.234 37.658 73.852 1.00 21.20 0 ATOM 4120 CA ILE 633 65.60838.788 73.014 1.00 19.86 0 ATOM 4121 CB ILE 633 66.085 38.308 71.6101.00 32.61 0 ATOM 4122 CG2 ILE 633 66.132 39.471 70.625 1.00 29.80 0ATOM 4123 CG1 ILE 633 67.490 37.716 71.700 1.00 28.84 0 ATOM 4124 CD1ILE 633 67.619 36.496 72.594 1.00 35.23 0 ATOM 4125 C ILE 633 64.55039.866 72.815 1.00 22.19 0 ATOM 4126 O ILE 633 64.864 41.053 72.878 1.0030.44 0 ATOM 4127 N TYR 634 63.303 39.473 72.588 1.00 32.62 0 ATOM 4129CA TYR 634 62.276 40.471 72.316 1.00 31.19 0 ATOM 4130 CB TYR 634 61.39540.006 71.147 1.00 13.98 0 ATOM 4131 CG TYR 634 62.233 39.755 69.9411.00 19.07 0 ATOM 4132 CD1 TYR 634 62.652 40.805 69.147 1.00 17.28 0ATOM 4133 CE1 TYR 634 63.498 40.587 68.074 1.00 16.13 0 ATOM 4134 CD2TYR 634 62.668 38.474 69.639 1.00 15.20 0 ATOM 4135 CE2 TYR 634 63.51538.236 68.571 1.00 18.14 0 ATOM 4136 CZ TYR 634 63.935 39.296 67.7861.00 21.41 0 ATOM 4137 OH TYR 634 64.789 39.069 66.720 1.00 20.11 0 ATOM4139 C TYR 634 61.430 41.025 73.446 1.00 28.63 0 ATOM 4140 O TYR 63460.380 41.637 73.188 1.00 19.20 0 ATOM 4141 N GLY 635 61.851 40.80774.690 1.00 38.67 0 ATOM 4143 CA GLY 635 61.114 41.407 75.786 1.00 39.390 ATOM 4144 C GLY 635 60.270 40.658 76.780 1.00 41.31 0 ATOM 4145 O GLY635 60.103 41.155 77.892 1.00 69.69 0 ATOM 4146 N PHE 636 59.725 39.50276.431 1.00 54.49 0 ATOM 4148 CA PHE 636 58.905 38.786 77.404 1.00 56.260 ATOM 4149 CB PHE 636 58.386 37.461 76.831 1.00 2.00 0 ATOM 4150 CG PHE636 57.275 36.845 77.639 1.00 2.00 0 ATOM 4151 CD1 PHE 636 56.293 37.63878.208 1.00 2.00 0 ATOM 4152 CD2 PHE 636 57.214 35.478 77.824 1.00 2.000 ATOM 4153 CE1 PHE 636 55.276 37.076 78.942 1.00 2.00 0 ATOM 4154 CE2PHE 636 56.210 34.919 78.549 1.00 2.00 0 ATOM 4155 CZ PHE 636 55.23835.716 79.111 1.00 2.00 0 ATOM 4156 C PHE 636 59.761 38.522 78.646 1.0055.84 0 ATOM 4157 O PHE 636 59.321 38.728 79.791 1.00 2.00 0 ATOM 4158 NTYR 637 61.005 38.114 78.415 1.00 2.00 0 ATOM 4160 CA TYR 637 61.91837.836 79.506 1.00 2.00 0 ATOM 4161 CB TYR 637 63.266 37.419 78.947 1.0018.03 0 ATOM 4162 CG TYR 637 64.345 37.308 79.986 1.00 13.39 0 ATOM 4163CD1 TYR 637 64.560 36.116 80.668 1.00 15.45 0 ATOM 4164 CE1 TYR 63765.563 36.007 81.603 1.00 14.13 0 ATOM 4165 CD2 TYR 637 65.163 38.39080.273 1.00 14.83 0 ATOM 4166 CE2 TYR 637 66.163 38.292 81.200 1.0013.98 0 ATOM 4167 CZ TYR 637 66.363 37.101 81.863 1.00 14.88 0 ATOM 4168OH TYR 637 67.379 37.015 82.784 1.00 13.10 0 ATOM 4170 C TYR 637 62.09139.056 80.411 1.00 2.00 0 ATOM 4171 O TYR 637 62.362 38.920 81.605 1.0020.03 0 ATOM 4172 N ASP 638 61.919 40.240 79.833 1.00 2.00 0 ATOM 4174CA ASP 638 62.086 41.484 80.554 1.00 2.00 0 ATOM 4175 CB ASP 638 62.70142.522 79.614 1.00 57.25 0 ATOM 4176 CG ASP 638 64.049 42.053 79.0461.00 66.80 0 ATOM 4177 OD1 ASP 638 64.078 41.539 77.905 1.00 65.06 0ATOM 4178 OD2 ASP 638 65.077 42.175 79.750 1.00 69.12 0 ATOM 4179 C ASP638 60.816 41.961 81.245 1.00 2.00 0 ATOM 4180 O ASP 638 60.884 42.53182.331 1.00 55.83 0 ATOM 4181 N GLU 639 59.656 41.724 80.644 1.00 2.00 0ATOM 4183 CA GLU 639 58.405 42.074 81.317 1.00 2.00 0 ATOM 4184 CB GLU639 67.210 41.774 60.419 1.00 64.74 0 ATOM 4185 CG GLU 639 57.051 42.72679.261 1.00 71.70 0 ATOM 4186 CD GLU 639 55.900 42.349 78.355 1.00 66.550 ATOM 4187 OE1 GLU 639 54.748 42.718 78.667 1.00 65.31 0 ATOM 4188 OE2GLU 639 56.152 41.684 77.329 1.00 72.66 0 ATOM 4189 C GLU 639 58.37241.145 82.547 1.00 2.00 0 ATOM 4190 O GLU 639 58.009 41.550 83.654 1.0065.95 0 ATOM 4191 N CYS 640 58.787 39.895 82.314 1.00 15.74 0 ATOM 4193CA CYS 640 58.859 38.843 83.323 1.00 15.74 0 ATOM 4194 CB CYS 640 59.18737.504 82.684 1.00 4.35 0 ATOM 4195 SG CYS 640 57.734 36.587 82.208 1.0013.44 0 ATOM 4196 C CYS 640 59.859 39.081 84.426 1.00 15.74 0 ATOM 4197O CYS 640 59.590 38.723 85.564 1.00 11.73 0 ATOM 4198 N LYS 641 61.02839.631 84.110 1.00 2.00 0 ATOM 4200 CA LYS 641 62.009 39.890 85.153 1.002.00 0 ATOM 4201 CB LYS 641 63.425 39.454 84.721 1.00 32.33 0 ATOM 4202CG LYS 641 64.225 40.436 83.869 1.00 32.55 0 ATOM 4203 CD LYS 641 65.73540.154 83.954 1.00 38.99 0 ATOM 4204 CE LYS 641 66.281 40.198 85.3861.00 41.14 0 ATOM 4205 NZ LYS 641 66.228 41.554 86.007 1.00 38.23 0 ATOM4209 C LYS 641 61.950 41.371 85.568 1.00 2.00 0 ATOM 4210 O LYS 64162.948 42.106 85.548 1.00 35.25 0 ATOM 4211 N ARG 642 60.750 41.80185.945 1.00 17.38 0 ATOM 4213 CA ARG 642 60.502 43.168 86.384 1.00 17.380 ATOM 4214 CB ARG 642 60.341 44.131 85.204 1.00 14.00 0 ATOM 4215 CGARG 642 61.636 44.613 84.561 1.00 23.15 0 ATOM 4216 CD ARG 642 61.46346.016 83.940 1.00 25.43 0 ATOM 4217 NE ARG 642 60.488 46.077 82.8461.00 33.87 0 ATOM 4219 CZ ARG 642 60.816 46.208 81.560 1.00 39.98 0 ATOM4220 NH1 ARG 642 62.097 46.291 81.199 1.00 44.40 0 ATOM 4223 NH2 ARG 64259.867 46.253 80.631 1.00 41.77 0 ATOM 4226 C ARG 642 59.200 43.09887.126 1.00 17.38 0 ATOM 4227 O ARG 642 59.114 43.455 88.288 1.00 26.010 ATOM 4228 N ARG 643 58.180 42.630 86.427 1.00 11.54 0 ATOM 4230 CA ARG643 56.869 42.500 87.018 1.00 11.54 0 ATOM 4231 CB ARG 643 55.804 42.39285.924 1.00 8.38 0 ATOM 4232 CG ARG 643 55.526 43.749 85.304 1.00 8.38 0ATOM 4233 CD ARG 643 54.377 43.745 84.341 1.00 8.38 0 ATOM 4234 NE ARG643 53.121 43.351 84.949 1.00 8.38 0 ATOM 4236 CZ ARG 643 51.939 43.56984.390 1.00 8.38 0 ATOM 4237 NH1 ARG 643 51.869 44.187 83.218 1.00 8.380 ATOM 4240 NH2 ARG 643 50.825 43.157 84.985 1.00 8.38 0 ATOM 4243 C ARG643 56.841 41.304 87.939 1.00 11.54 0 ATOM 4244 O ARG 643 56.156 41.30988.969 1.00 8.38 0 ATOM 4245 N TYR 644 57.606 40.285 87.565 1.00 2.00 0ATOM 4247 CA TYR 644 57.703 39.052 88.333 1.00 2.00 0 ATOM 4248 CB TYR644 56.784 37.972 87.741 1.00 14.67 0 ATOM 4249 CG TYR 644 55.338 38.37887.740 1.00 14.67 0 ATOM 4250 CD1 TYR 644 54.695 38.723 86.554 1.0014.67 0 ATOM 4251 CE1 TYR 644 53.359 39.140 86.54B 1.00 14.67 0 ATOM4252 CD2 TYR 644 54.617 38.453 88.922 1.00 14.67 0 ATOM 4253 CE2 TYR 64453.289 38.866 88.929 1.00 14.67 0 ATOM 4254 CZ TYR 644 52.665 39.20987.739 1.00 14.67 0 ATOM 4255 OH TYR 644 51.349 39.629 87.741 1.00 14.670 ATOM 4257 C TYR 644 59.160 38.598 88.330 1.00 2.00 0 ATOM 4258 O TYR644 60.062 39.412 88.575 1.00 14.67 0 ATOM 4259 N ASN 645 59.399 37.32188.028 1.00 2.00 0 ATOM 4261 CA ASN 645 60.753 36.794 88.021 1.00 2.00 0ATOM 4262 CB ASN 645 61.103 36.147 89.379 1.00 16.19 0 ATOM 4263 CG ASN645 60.081 35.097 89.846 1.00 16.19 0 ATOM 4264 OD1 ASN 645 59.46634.378 89.054 1.00 16.19 0 ATOM 4265 ND2 ASN 645 59.916 35.009 91.1501.00 16.19 0 ATOM 4268 C ASN 645 61.051 35.809 86.927 1.00 2.00 0 ATOM4269 O ASN 645 60.156 35.217 86.333 1.00 16.19 0 ATOM 4270 N ILE 64662.342 35.648 86.678 1.00 2.00 0 ATOM 4272 CA ILE 646 62.856 34.71685.689 1.00 2.00 0 ATOM 4273 CB ILE 646 64.390 34.593 85.830 1.00 2.81 0ATOM 4274 CG2 ILE 646 64.917 33.316 85.196 1.00 2.59 0 ATOM 4275 CG1 ILE646 65.054 35.814 85.221 l.00 2.59 0 ATOM 4276 CD1 ILE 646 66.492 35.95585.666 1.00 8.72 0 ATOM 4277 C ILE 646 62.214 33.339 85.886 1.00 2.00 0ATOM 4278 O ILE 646 61.875 32.675 84.915 1.00 6.42 0 ATOM 4279 N LYS 64762.031 32.918 87.137 1.00 2.00 0 ATOM 4281 CA LYS 647 61.441 31.61487.411 1.00 2.00 0 ATOM 4282 CB LYS 647 61.347 31.387 88.920 1.00 79.290 ATOM 4283 CG LYS 647 62.714 31.450 89.594 1.00 84.06 0 ATOM 4284 CDLYS 647 63.727 30.622 88.804 1.00 88.57 0 ATOM 4285 CE LYS 647 61.15731.090 89.028 1.00 86.67 0 ATDM 4286 NZ LYS 647 66.029 30.668 87.8861.00 91.44 0 ATOM 4290 C LYS 647 60.083 31.478 86.738 1.00 2.00 0 ATOM4291 O LYS 647 59.751 30.426 86.187 1.00 74.01 0 ATOM 4292 N LEU 64859.322 32.563 86.763 1.00 25.02 0 ATOM 4294 CA LEU 648 58.012 32.59986.136 1.00 23.81 0 ATOM 4295 CB LEU 648 57.300 33.918 86.460 1.00 2.000 ATOM 4296 CG LEU 648 55.802 33.929 86.178 1.00 2.00 0 ATOM 4297 CD1LEU 648 51.145 32.777 86.951 1.00 2.00 0 ATOM 4298 CD2 LEU 648 55.20635.262 86.574 1.00 2.00 0 ATOM 4299 C LEU 648 58.245 32.495 84.637 1.0028.16 0 ATOM 4300 O LEU 648 57.562 31.751 83.934 1.00 2.00 0 ATOM 4301 NTRP 649 59.227 33.243 84.151 1.00 42.46 0 ATOM 4303 CA TRP 649 59.55433.224 82.738 1.00 40.96 0 ATOM 4304 CB TRP 649 60.719 34.161 82.4651.00 11.77 0 ATOM 4305 CG TRP 649 61.206 34.051 81.081 1.00 14.98 0 ATOM4306 CD2 TRP 649 62.393 33.394 80.653 1.00 14.05 0 ATOM 4307 CE2 TRP 64962.460 33.527 79.246 1.00 12.39 0 ATOM 4308 CE3 TRP 649 63.412 32.70381.320 1.00 25.06 0 ATOM 4309 CD1 TRP 649 60.607 34.546 79.951 1.0017.81 0 ATOM 4310 NE1 TRP 649 61.356 34.232 78.846 1.00 15.30 0 ATOM4312 CZ2 TRP 649 63.510 32.995 78.495 1.00 13.58 0 ATOM 4313 CZ3 TRP 64964.456 32.174 80.576 1.00 15.93 0 ATOM 4314 CH2 TRP 649 64.497 32.32379.175 1.00 16.10 0 ATOM 4315 C TRP 649 59.900 31.802 82.292 1.00 40.420 ATOM 4316 O TRP 649 59.392 31.315 81.285 1.00 12.84 0 ATOM 4317 N LYS650 60.748 31.138 83.069 1.00 2.00 0 ATOM 4319 CA LYS 650 61.179 29.77682.796 1.00 2.00 0 ATOM 4320 CB LYS 650 62.257 29.371 93.795 1.00 19.180 ATOM 4321 CG LYS 650 63.420 30.343 83.820 1.00 4.52 0 ATOM 4322 CD LYS650 64.541 29.898 84.718 1.00 3.83 0 ATOM 4323 CE LYS 650 65.148 28.59384.221 1.00 11.09 0 ATOM 4324 NZ LYS 650 66.096 27.974 85.202 1.00 11.030 ATOM 4328 C LYS 650 59.976 28.851 82.892 1.00 2.00 0 ATOM 4329 O LYS650 59.926 27.805 82.239 1.00 5.16 0 ATOM 4330 N THR 651 58.997 29.23483.702 1.00 10.53 0 ATOM 4332 CA THR 651 57.792 28.422 83.838 1.00 14.100 ATOM 4333 CB THR 651 56.915 28.882 85.038 1.00 23.55 0 ATOM 4334 OG1THR 651 57.613 28.622 86.268 1.00 23.26 0 ATOM 4336 CG2 THR 651 55.58228.150 85.047 1.00 24.53 0 ATOM 4337 C THR 651 57.012 28.543 82.539 1.0014.03 0 ATOM 4338 O THR 651 56.599 27.534 81.951 1.00 23.01 0 ATOM 4339N PHE 652 56.837 29.779 82.082 1.00 2.00 0 ATOM 4341 CA PHE 652 56.12730.032 80.840 1.00 2.00 0 ATOM 4342 CB PHE 652 56.225 31.511 80.440 1.002.00 0 ATOM 4343 CG PHE 652 55.027 32.320 80.821 1.00 2.00 0 ATOM 4344CD1 PHE 652 55.164 33.486 81.536 1.00 2.00 0 ATOM 4345 CD2 PHE 65253.753 31.919 80.459 1.00 2.00 0 ATOM 4346 CE1 PHE 652 54.039 34.24581.884 1.00 2.00 0 ATOM 4347 CE2 PHE 652 52.630 32.678 80.808 1.00 2.000 ATOM 4348 CZ PHE 652 52.775 33.832 81.515 1.00 2.00 0 ATOM 4349 C PHE652 56.717 29.160 79.743 1.00 2.00 0 ATOM 4350 O PHE 652 55.981 28.45679.067 1.00 2.00 0 ATOM 4351 N THR 653 58.039 29.143 79.608 1.00 2.00 0ATOM 4353 CA THR 653 58.631 28.344 78.552 1.00 2.00 0 ATOM 4354 CB THR653 60.126 28.538 78.429 1.00 2.14 0 ATOM 4355 OG1 THR 653 60.804 27.59279.255 1.00 2.14 0 ATOM 4357 CG2 THR 653 60.499 29.945 78.787 1.00 2.140 ATOM 4358 C THR 653 58.371 26.846 78.588 1.00 2.00 0 ATOM 4359 O THR653 58.452 26.196 77.555 1.00 4.20 0 ATOM 4360 N ASP 654 58.064 26.27179.743 1.00 2.00 0 ATOM 4362 CA ASP 654 57.803 24.839 79.749 1.00 2.00 0ATOM 4363 CB ASP 654 58.083 24.225 81.119 1.00 25.83 0 ATOM 4364 CG ASP654 58.607 22.794 81.018 1.00 26.00 0 ATOM 4365 CD1 ASP 654 59.16322.420 79.960 1.00 29.22 0 ATOM 4366 CD2 ASP 654 58.472 22.043 82.0061.00 28.12 0 ATOM 4367 C ASP 654 56.367 24.603 79.332 1.00 2.00 0 ATOM4368 O ASP 654 56.014 23.518 78.874 1.00 18.75 0 ATOM 4369 N CYS 65555.537 25.625 79.503 1.00 27.58 0 ATOM 4371 CA CYS 655 54.146 25.54679.095 1.00 27.58 0 ATOM 4372 CB CYS 655 53.333 26.675 79.722 1.00 8.450 ATOM 4373 SG CYS 655 51.756 26.978 78.901 1.00 8.45 0 ATOM 4374 C CYS655 54.162 25.683 77.575 1.00 27.58 0 ATOM 4375 O CYS 655 53.565 24.86376.865 1.00 8.45 0 ATOM 4376 N PHE 656 54.871 26.709 77.088 1.00 7.64 0ATOM 4378 CA PHE 656 55.018 26.966 75.653 1.00 7.64 0 ATOM 4379 CB PHE656 55.967 28.145 75.402 1.00 12.44 0 ATOM 4380 CG PHE 656 55.384 29.48375.747 1.00 12.44 0 ATOM 4381 CD1 PHE 656 54.073 29.591 76.242 1.0012.44 0 ATOM 4382 CD2 PHE 656 56.144 30.641 75.594 1.00 12.44 0 ATOM4383 CE1 PHE 656 53.525 30.840 76.585 1.00 12.44 0 ATOM 4384 CE2 PHE 65655.614 31.891 75.929 1.00 12.44 0 ATOM 4385 CZ PHE 656 54.296 31.99076.430 1.00 12.44 0 ATOM 4386 C PHE 656 55.564 25.705 74.961 1.00 7.64 0ATOM 4387 O PHE 656 55.033 25.274 73.940 1.00 12.44 0 ATOM 4388 N ASN657 56.595 25.099 75.543 1.00 2.00 0 ATOM 4390 CA ASN 657 57.198 23.89675.006 1.00 2.00 0 ATOM 4391 CB ASN 657 58.353 23.425 75.892 1.00 12.640 ATOM 4392 CG ASN 657 59.614 24.233 75.690 1.00 12.64 0 ATOM 4393 OD1ASN 657 59.564 25.404 75.350 1.00 12.64 0 ATOM 4394 ND2 ASN 657 60.75723.607 75.896 1.00 12.64 0 ATOM 4397 C ASN 657 56.202 22.772 74.881 1.002.00 0 ATOM 4398 O ASN 657 56.585 21.668 74.532 1.00 12.64 0 ATOM 4399 NCYS 658 54.934 23.017 75.196 1.00 2.00 0 ATOM 4401 CA CYS 658 53.93021.964 75.080 1.00 2.00 0 ATOM 4402 CB CYS 658 53.543 21.468 76.475 1.0010.87 0 ATOM 4403 SG CYS 658 55.004 20.919 77.397 1.00 10.87 0 ATOM 4404C CYS 658 52.708 22.390 74.258 1.00 2.00 0 ATOM 4405 O CYS 658 51.72621.659 74.162 1.00 10.87 0 ATOM 4406 N LEU 659 52.799 23.560 73.634 1.0011.17 0 ATOM 4408 CA LEU 659 51.739 24.085 72.781 1.00 11.17 0 ATOM 4409CB LEU 659 51.998 25.569 72.470 1.00 2.00 0 ATOM 4410 CG LEU 659 51.75726.629 73.542 1.00 2.00 0 ATOM 4411 CD1 LEU 659 52.286 27.965 73.1231.00 2.00 0 ATOM 4412 CD2 LEU 659 50.307 26.759 73.767 1.00 2.00 0 ATOM4413 C LEU 659 51.651 23.298 71.450 1.00 11.17 0 ATOM 4414 O LEU 65952.681 22.845 70.900 1.00 2.00 0 ATOM 4415 N PRO 660 50.415 23.10970.931 1.00 14.96 0 ATOM 4416 CD PRO 660 49.134 23.535 71.523 1.00 2.000 ATOM 4417 CA PRO 660 50.166 22.398 69.680 1.00 14.96 0 ATOM 4418 CBPRO 660 48.640 22.381 69.590 1.00 2.00 0 ATOM 4419 CG PRO 660 48.19422.472 71.028 1.00 2.00 0 ATOM 4420 C PRO 660 50.796 23.248 68.579 1.0014.96 0 ATOM 4421 O PRO 660 50.888 24.474 68.709 1.00 2.00 0 ATOM 4422 NILE 661 51.195 22.603 67.488 1.00 28.66 0 ATOM 4424 CA ILE 661 51.88023.279 66.397 1.00 30.55 0 ATOM 4425 CB ILE 661 53.126 22.480 66.0361.00 19.83 0 ATOM 4426 CG2 ILE 661 54.064 22.432 67.228 1.00 22.82 0ATOM 4427 CG1 ILE 661 52.728 21.050 65.669 1.00 24.89 0 ATOM 4428 CD1ILE 661 53.896 20.161 65.299 1.00 29.65 0 ATOM 4429 C ILE 661 51.07423.577 65.132 1.00 29.24 0 ATOM 4430 O ILE 661 51.428 24.493 64.372 1.0020.63 0 ATOM 4431 N ALA 662 50.004 22.810 64.915 1.00 22.79 0 ATOM 4433CA ALA 662 49.112 22.970 63.751 1.00 22.79 0 ATOM 4434 CB ALA 662 49.65222.187 62.529 1.00 2.00 0 ATOM 4435 C ALA 662 47.683 22.507 64.086 1.0022.79 0 ATOM 4436 O ALA 662 47.439 21.887 65.139 1.00 2.00 0 ATOM 4437 NALA 663 46.739 22.816 63.205 1.00 2.00 0 ATOM 4439 CA ALA 663 45.34722.438 63.410 1.00 2.00 0 ATOM 4440 CB ALA 663 44.599 23.535 64.124 1.0018.31 0 ATOM 4441 C ALA 663 44.748 22.223 62.053 1.00 2.00 0 ATOM 4442 OALA 663 45.323 22.623 61.041 1.00 22.14 0 ATOM 4443 N ILE 664 43.60021.565 62.028 1.00 16.69 0 ATOM 4445 CA ILE 664 42.892 21.300 60.7881.00 16.69 0 ATOM 4446 CB ILE 664 43.240 19.885 60.208 1.00 9.81 0 ATOM4447 CG2 ILE 664 42.396 19.597 58.976 1.00 9.81 0 ATOM 4448 CG1 ILE 66444.724 19.810 59.819 1.00 9.81 0 ATOM 4449 CD1 ILE 664 45.161 18.45059.316 1.00 9.81 0 ATOM 4450 C ILE 664 41.408 21.397 61.118 1.00 16.69 0ATOM 4451 O ILE 664 40.880 20.603 61.899 1.00 9.81 0 ATOM 4452 N VAL 66540.754 22.408 60.561 1.00 14.09 0 ATOM 4454 CA VAL 665 39.332 22.60860.777 1.00 14.09 0 ATOM 4455 CB VAL 665 38.954 24.091 60.742 1.00 2.000 ATOM 4456 CG1 VAL 665 37.450 24.239 60.710 1.00 2.00 0 ATOM 4457 CG2VAL 665 38.500 24.788 61.950 1.00 2.00 0 ATOM 4458 C VAL 665 38.50921.873 59.729 1.00 14.09 0 ATOM 4459 O VAL 665 38.768 21.972 58.524 1.002.00 0 ATOM 4460 N ASP 666 37.517 21.137 60.219 1.00 2.00 0 ATOM 4462 CAASP 666 36.595 20.345 59.407 1.00 2.00 0 ATOM 4463 CB ASP 666 35.44321.235 58.934 1.00 65.64 0 ATOM 4464 CG ASP 666 34.545 21.675 60.0811.00 73.26 0 ATOM 4465 OD1 ASP 666 33.763 20.838 60.578 1.00 71.65 0ATOM 4466 OD2 ASP 666 34.623 22.851 60.493 1.00 75.52 0 ATOM 4467 C ASP666 37.234 19.577 58.235 1.00 2.00 0 ATOM 4468 O ASP 666 36.648 19.46757.158 1.00 57.66 0 ATOM 4469 N GLU 667 38.439 19.048 58.472 1.00 17.200 ATOM 4471 CZ GLU 667 39.203 18.270 57.489 1.00 17.69 0 ATOM 4472 CBGLU 667 38.455 16.981 57.138 1.00 42.87 0 ATOM 4473 CG GLU 667 38.17016.101 58.345 1.00 52.20 0 ATOM 4474 CD GLU 667 37.457 14.806 57.9881.00 53.85 0 ATOM 4475 OB1 GLU 667 36.222 14.717 58.211 1.00 51.15 0ATOM 4476 OB2 GLU 667 38.139 13.877 57.494 1.00 55.67 0 ATOM 4477 C GLU667 39.584 19.015 56.207 1.00 17.24 0 ATOM 4478 O GLU 667 40.146 18.42155.286 1.00 34.30 0 ATOM 4479 N LYS 668 39.314 20.319 56.172 1.00 26.530 ATOM 4481 CA LYS 668 39.615 21.133 55.002 1.00 20.22 0 ATOM 4482 CBLYS 668 38.318 21.709 54.410 1.00 13.48 0 ATOM 4483 CC LYS 668 37.38320.628 53.859 1.00 13.48 0 ATOM 4484 CD LYS 668 38.119 19.779 52.8191.00 13.48 0 ATOM 4485 CE LYS 668 37.341 18.551 52.398 1.00 16.60 0 ATOM4486 NZ LYS 668 36.247 17.564 51.739 1.00 18.66 0 ATOM 4490 C LYS 66840.636 22.244 55.240 1.00 19.61 0 ATOM 4491 O LYS 668 41.676 22.26454.584 1.00 13.48 0 ATOM 4492 N ILE 669 40.356 23.166 56.158 1.00 2.00 0ATOM 4494 CA ILE 669 41.295 24.263 56.424 1.00 2.00 0 ATOM 4495 CB ILE669 40.617 25.456 57.183 1.00 2.00 0 ATOM 4496 CG2 ILE 669 41.521 26.68057.166 1.00 2.00 0 ATOM 4497 CG1 ILE 669 39.298 25.839 56.518 1.00 2.000 ATOM 4498 CD1 ILE 669 38.581 26.960 57.189 1.00 2.00 0 ATOM 4499 C ILE669 42.439 23.721 57.279 1.00 2.00 0 ATOM 4500 O ILE 669 42.201 23.03458.269 1.00 2.00 0 ATOM 4501 N PHE 670 43.673 24.010 56.892 1.00 2.00 0ATOM 4503 CA PHE 670 44.841 23.551 57.641 1.00 2.00 0 ATOM 4504 CB PHE670 45.804 22.801 56.718 1.00 2.00 0 ATOM 4505 CG PHE 670 47.182 22.61457.291 1.00 2.00 0 ATOM 4506 CD1 PHE 670 47.503 21.473 58.007 1.00 2.000 ATOM 4507 CD2 PHE 670 48.163 23.569 57.093 1.00 2.00 0 ATOM 4508 CE1PHE 670 48.769 21.288 58.506 1.00 2.00 0 ATOM 4509 CE2 PHE 670 49.43623.384 57.596 1.00 2.00 0 ATOM 4510 CE PHE 670 49.737 22.241 58.302 1.002.00 0 ATOM 4511 C PHE 670 45.528 24.780 58.190 1.00 2.00 0 ATOM 4512 OPHE 670 45.901 25.652 57.421 1.00 2.00 0 ATOM 4513 N CYS 671 45.72524.839 59.501 1.00 2.00 0 ATOM 4515 CA CYS 671 46.353 25.992 60.120 1.002.00 0 ATOM 4516 CB CYS 671 45.395 26.596 61.136 1.00 14.45 0 ATOM 4517SG CYS 671 43.708 26.779 60.568 1.00 25.33 0 ATOM 4518 C CYS 671 47.68525.701 60.816 1.00 2.00 0 ATOM 4519 O CYS 671 47.921 24.589 61.310 1.008.01 0 ATOM 4520 N CYS 672 48.546 26.714 60.845 1.00 2.00 0 ATOM 4522 CACYS 672 49.845 26.666 61.515 1.00 2.00 0 ATOM 4523 CB CYS 672 50.82625.741 60.792 1.00 9.00 0 ATOM 4524 SG CYS 672 51.494 26.372 59.265 1.0011.24 0 ATOM 4525 C CYS 672 50.336 28.122 61.525 1.00 2.00 0 ATOM 4526 OCYS 672 49.751 28.963 60.850 1.00 9.00 0 ATOM 4527 N HIS 673 51.36928.440 62.300 1.00 17.94 0 ATOM 4529 CA HIS 673 51.855 29.817 62.3601.00 17.94 0 ATOM 4530 C HIS 673 52.456 30.359 61.065 1.00 17.94 0 ATOM4531 O HIS 673 51.960 31.349 60.514 1.00 2.00 0 ATOM 4532 CB HIS 67352.894 29.965 63.459 1.00 2.00 0 ATOM 4533 CG HIS 673 53.283 31.38363.724 1.00 2.00 0 ATOM 4534 ND1 HIS 673 52.388 32.377 64.033 1.00 2.000 ATOM 4536 CD2 HIS 673 54.503 31.975 63.718 1.00 2.00 0 ATOM 4537 NE2HIS 673 54.371 33.330 64.019 1.00 2.00 0 ATOM 4538 CE1 HIS 673 53.07233.512 64.199 1.00 2.00 0 ATOM 4539 N GLY 674 53.545 29.721 60.626 1.002.00 0 ATOM 4541 CA GLY 674 54.260 30.101 59.417 1.00 2.00 0 ATOM 4542 CGLY 674 53.773 29.407 58.163 1.00 2.00 0 ATOM 4543 O GLY 674 58.41030.063 57.203 1.00 11.62 0 ATOM 4544 N GLY 675 53.759 28.087 58.136 1.006.25 0 ATOM 4546 CA GLY 675 53.286 27.440 56.931 1.00 6.25 0 ATOM 4547 CGLY 675 53.823 26.061 56.604 1.00 6.25 0 ATOM 4548 O GLY 675 53.86725.173 57.452 1.00 28.15 0 ATOM 4549 N LEU 676 54.241 25.883 55.356 1.002.00 0 ATOM 4551 CA LEU 676 54.723 24.591 54.886 1.00 2.00 0 ATOM 4552CB LEU 676 54.336 24.409 53.418 1.00 2.00 0 ATOM 4553 CG LEU 676 52.82724.625 53.244 1.00 2.00 0 ATOM 4554 CD1 LEU 676 52.412 24.556 51.7771.00 2.00 0 ATOM 4555 CD2 LEU 676 52.100 23.575 54.075 1.00 2.00 0 ATOM4556 C LEU 676 56.207 24.333 55.095 1.00 2.00 0 ATOM 4557 O LEU 67656.981 25.248 55.381 1.00 2.00 0 ATOM 4558 N SER 677 56.582 23.06954.945 1.00 12.63 0 ATOM 4560 CA SER 677 57.946 22.617 55.141 1.00 12.630 ATOM 4561 CB SER 677 58.083 21.985 56.539 1.00 2.43 0 ATOM 4562 OG SER677 59.248 21.192 56.649 1.00 2.03 0 ATOM 4564 C SER 677 58.247 21.55654.095 1.00 12.63 0 ATOM 4565 O SER 677 57.405 20.684 53.838 1.00 10.720 ATOM 4566 N PRO 678 59.449 21.605 53.484 1.00 2.00 0 ATOM 4567 CD PRO678 60.474 22.638 53.681 1.00 15.59 0 ATOM 4568 CA PRO 678 59.887 20.64752.469 1.00 2.00 0 ATOM 4569 CB PRO 678 61.329 21.076 52.184 1.00 15.590 ATOM 4570 CG PRO 678 61.299 22.517 52.416 1.00 15.59 0 ATOM 4571 C PRO678 59.859 19.234 53.032 1.00 2.00 0 ATOM 4572 O PRO 678 59.990 18.26952.293 1.00 15.59 0 ATOM 4573 N ASP 679 59.685 19.112 54.342 1.00 2.00 0ATOM 4575 CA ASP 679 59.687 17.813 54.992 1.00 2.00 0 ATOM 4576 CB ASP679 60.510 17.926 56.271 1.00 26.28 0 ATOM 4577 CG ASP 679 61.766 18.75856.073 1.00 24.05 0 ATOM 4578 CD1 ASP 679 62.687 18.269 55.383 1.0032.92 0 ATOM 4579 CD2 ASP 679 61.826 19.902 56.586 1.00 28.53 0 ATOM4580 C ASP 679 58.303 17.278 55.314 1.00 2.00 0 ATOM 4581 O ASP 67958.129 16.094 55.576 1.00 24.04 0 ATOM 4582 N LEU 680 57.315 18.14955.279 1.00 2.00 0 ATOM 4584 CA LEU 680 55.970 17.747 55.622 1.00 2.00 0ATOM 4585 CB LEU 680 55.147 19.000 55.916 1.00 4.45 0 ATOM 4586 CG LEU680 53.683 18.788 56.262 1.00 2.86 0 ATOM 4587 CD1 LEU 680 53.540 17.79157.398 1.00 2.86 0 ATOM 4588 CD2 LEU 680 53.085 20.134 56.582 1.00 2.860 ATOM 4589 C LEU 680 55.262 16.862 54.588 1.00 2.00 0 ATOM 4590 O LEU680 54.421 17.331 53.814 1.00 15.95 0 ATOM 4591 N GLN 681 55.588 15.57854.561 1.00 2.00 0 ATOM 4593 CA GLN 681 54.922 14.686 53.611 1.00 2.00 0ATOM 4594 CB GLN 681 55.719 13.410 53.885 1.00 36.99 0 ATOM 4595 CG GLN681 57.097 13.620 52.863 1.00 36.99 0 ATOM 4596 CD GLN 681 57.716 12.32152.468 1.00 36.99 0 ATOM 4597 CE1 GLN 681 57.589 11.894 51.326 1.0036.99 0 ATOM 4598 NE2 GLN 681 58.377 11.663 53.409 1.00 36.99 0 ATOM4601 C GLN 681 53.551 14.319 54.159 1.00 2.00 0 ATOM 4602 O GLN 68152.547 14.392 53.445 1.00 11.55 0 ATOM 4603 N SER 682 53.505 13.92355.423 1.00 39.00 0 ATOM 4605 CA SER 682 52.238 13.566 56.027 1.00 42.030 ATOM 4606 CB SER 682 52.131 12.048 56.224 1.00 2.00 0 ATOM 4607 CG SER682 53.198 11.507 56.993 1.00 2.00 0 ATOM 4609 C SER 682 52.003 14.28457.343 1.00 38.75 0 ATOM 4610 O SER 682 52.941 14.742 58.009 1.00 2.00 0ATOM 4611 N MET 683 50.730 14.397 57.698 1.00 15.16 0 ATOM 4613 CA MET683 50.338 15.029 58.938 1.00 15.16 0 ATOM 4614 CB MET 683 48.820 15.06959.044 1.00 11.68 0 ATOM 4615 CG MET 683 48.099 15.565 57.799 1.00 12.750 ATOM 4616 SD MET 683 48.197 17.328 57.477 1.00 11.68 0 ATOM 4617 CEMET 683 49.486 17.429 56.200 1.00 12.61 0 ATOM 4618 C MET 683 50.90614.131 60.024 1.00 15.16 0 ATOM 4619 O MET 683 51.215 14.585 61.116 1.0015.01 0 ATOM 4620 N GLU 684 51.050 12.848 59.705 1.00 40.22 0 ATOM 4622CA GLU 684 51.597 11.881 60.644 1.00 41.85 0 ATOM 4623 CB GLU 684 52.00710.594 59.937 1.00 63.04 0 ATOM 4624 CG GLU 684 52.850 9.689 60.821 1.0072.94 0 ATOM 4625 CD GLU 684 53.111 8.343 60.206 1.00 77.43 0 ATOM 4626OE1 GLU 684 52.139 7.716 59.730 1.00 77.96 0 ATOM 4627 OE2 GLU 68454.285 7.909 60.201 1.00 79.67 0 ATOM 4628 C GLU 684 52.819 12.47161.297 1.00 40.42 0 ATOM 4629 O GLU 684 52.932 12.495 62.517 1.00 62.000 ATOM 4630 N GLN 685 53.727 12.959 60.466 1.00 2.00 0 ATOM 4632 CA GLN685 54.945 13.559 60.952 1.00 2.00 0 ATOM 4633 CB GLN 685 55.703 14.20059.797 1.00 52.45 0 ATOM 4634 CG GLN 685 56.162 13.188 58.775 1.00 57.010 ATOM 4635 CD GLN 685 56.779 13.832 57.574 1.00 58.68 0 ATOM 4636 OE1GLN 685 56.258 13.728 56.471 1.00 68.68 0 ATOM 4637 NE2 GLN 685 57.89514.510 57.778 1.00 65.77 0 ATOM 4640 C GLN 685 54.627 14.585 62.029 1.002.00 0 ATOM 4641 O GLN 685 55.312 14.631 63.056 1.00 54.77 0 ATOM 4642 NILE 686 53.579 15.387 61.816 1.00 16.80 0 ATOM 4644 CA ILE 686 53.17916.391 62.807 1.00 14.42 0 ATOM 4645 CB ILE 686 51.990 17.270 62.3071.00 2.00 0 ATOM 4646 CG2 ILE 686 51.754 18.441 63.247 1.00 2.00 0 ATOM4647 CG1 ILE 686 52.304 17.869 60.944 1.00 2.00 0 ATOM 4648 CD1 ILE 68651.139 18.668 60.369 1.00 2.00 0 ATOM 4649 C ILE 686 52.761 15.63364.079 1.00 13.83 0 ATOM 4650 O ILE 686 53.289 15.884 65.165 1.00 2.00 0ATOM 4651 N ARG 687 51.856 14.669 63.914 1.00 3.14 0 ATOM 4653 CA ARG687 51.367 13.855 65.027 1.00 3.14 0 ATOM 4654 CB ARG 687 50.307 12.83564.564 1.00 23.50 0 ATOM 4655 CG ARG 687 49.266 13.324 63.559 1.00 26.960 ATOM 4656 CD ARG 687 48.288 12.211 63.083 1.00 35.95 0 ATOM 4657 NEARG 687 48.878 11.199 62.190 1.00 41.22 0 ATOM 4659 CZ ARG 687 49.58310.138 62.594 1.00 45.24 0 ATOM 4660 NH1 ARG 687 50.062 9.283 61.7021.00 43.65 0 ATOM 4663 NH2 ARG 687 49.827 9.922 63.887 1.00 36.26 0 ATOM4666 C ARG 687 52.504 13.060 65.665 1.00 3.14 0 ATOM 4667 O ARG 68752.260 12.327 66.613 1.00 18.08 0 ATOM 4668 N ARG 688 53.722 13.15765.136 1.00 11.08 0 ATOM 4670 CA ARG 688 54.841 12.402 65.694 1.00 10.970 ATOM 4671 CB ARG 688 55.576 11.624 64.595 1.00 52.72 0 ATOM 4672 CGARG 688 54.794 10.485 63.995 1.00 52.20 0 ATOM 4673 CD ARG 688 54.4219.463 65.035 1.00 55.85 0 ATOM 4674 NE ARG 688 53.634 8.372 64.468 1.0050.50 0 ATOM 4676 CZ ARG 688 54.145 7.254 63.956 1.00 54.57 0 ATOM 4677NH1 ARG 688 53.335 6.326 63.465 1.00 53.02 0 ATOM 4680 NH2 ARG 68855.457 7.056 63.931 1.00 50.07 0 ATOM 4683 C ARG 688 55.853 13.25766.457 1.00 9.37 0 ATOM 4684 O ARG 688 56.771 12.719 67.084 1.00 54.51 0ATOM 4685 N ILE 689 55.698 14.576 66.398 1.00 38.26 0 ATOM 4687 CA ILE689 56.614 15.487 67.081 1.00 36.62 0 ATOM 4688 CB ILE 689 56.319 16.95266.673 1.00 2.00 0 ATOM 4689 CG2 ILE 689 57.248 17.901 67.399 1.00 2.000 ATOM 4690 CG1 ILE 689 56.493 17.119 65.160 1.00 2.00 0 ATOM 4691 CD1ILE 689 56.670 18.556 64.711 1.00 2.00 0 ATOM 4692 C ILE 689 56.55215.347 68.620 1.00 42.28 0 ATOM 4693 O ILE 689 55.468 15.462 69.226 1.002.00 0 ATOM 4694 N MET 690 57.710 15.084 69.242 1.00 2.00 0 ATOM 4696 CAMET 690 57.794 14.930 70.704 1.00 2.00 0 ATOM 4697 CB MET 690 59.20414.516 71.138 1.00 31.54 0 ATOM 4698 CG MET 690 59.657 13.160 70.6161.00 36.54 0 ATOM 4699 SD MET 690 58.703 11.742 71.207 1.00 43.14 0 ATOM4700 CE MET 690 59.784 10.411 70.721 1.00 40.70 0 ATOM 4701 C MET 69057.491 16.313 71.227 1.00 2.00 0 ATOM 4702 O MET 690 58.189 17.26970.864 1.00 21.20 0 ATOM 4703 N ARG 691 56.489 16.441 72.092 1.00 59.620 ATOM 4705 CA ARG 691 56.135 17.780 72.507 1.00 65.12 0 ATOM 4706 CBARG 691 54.678 17.879 72.869 1.00 2.00 0 ATOM 4707 CG ARG 691 54.07719.116 72.206 1.00 2.00 0 ATOM 4708 CD ARG 691 52.598 19.166 72.384 1.002.00 0 ATOM 4709 NE ARG 691 52.082 17.820 72.551 1.00 2.00 0 ATOM 4711CZ ARG 691 50.832 17.540 72.862 1.00 2.00 0 ATOM 4712 NH1 ARG 691 49.95718.525 71.024 1.00 2.00 0 ATOM 4715 NH2 ARG 691 50.482 16.273 73.0461.00 2.00 0 ATOM 4718 C ARG 691 56.930 18.632 73.458 1.00 64.88 0 ATOM4719 O ARG 691 57.176 19.794 73.100 1.00 2.00 0 ATOM 4720 N PRO 69257.265 18.151 74.693 1.00 0.89 0 ATOM 4721 CD PRO 692 56.930 16.94775.482 1.00 19.88 0 ATOM 4722 CA PRO 692 58.060 19.115 75.500 1.00 0.770 ATOM 4723 CB PRO 692 58.305 18.357 76.811 1.00 21.87 0 ATOM 4724 CGPRO 692 57.071 17.462 76.911 1.00 19.02 0 ATOM 4725 C PRO 692 59.32719.317 74.643 1.00 0.10 0 ATOM 4726 O PRO 692 60.258 18.502 74.690 1.0022.15 0 ATOM 4727 N THR 693 59.310 20.362 73.811 1.00 2.00 0 ATOM 4729CA THR 693 60.395 20.605 72.896 1.00 2.00 0 ATOM 4730 CB THR 693 60.15319.842 71.569 1.00 39.91 0 ATOM 4731 OG1 THR 693 61.310 19.947 70.7281.00 47.46 0 ATOM 4733 CG2 THR 693 58.944 20.419 70.830 1.00 46.31 0ATOM 4734 C THR 693 60.567 22.057 72.560 1.00 2.00 0 ATOM 4735 O THR 69359.640 22.848 72.678 1.00 41.07 0 ATOM 4736 N ASP 694 61.782 22.39972.154 1.00 4.51 0 ATOM 4738 CA ASP 694 62.075 23.747 71.736 1.00 4.51 0ATOM 4739 CB ASP 694 63.429 24.203 72.283 1.00 83.74 0 ATOM 4740 CG ASP694 63.337 25.520 73.041 1.00 83.74 0 ATOM 4741 OD1 ASP 694 63.23125.484 74.285 1.00 83.74 0 ATOM 4742 OD2 ASP 694 63.366 26.594 72.4001.00 83.74 0 ATOM 4743 C ASP 694 62.101 23.682 70.201 1.00 4.51 0 ATOM4744 O ASP 694 62.403 22.632 69.629 1.00 83.74 0 ATOM 4745 N VAL 69561.743 24.784 69.546 1.00 31.71 0 ATOM 4747 CA VAL 695 61.760 24.86468.087 1.00 37.86 0 ATOM 4748 CB VAL 695 61.212 26.224 67.623 1.00 72.520 ATOM 4749 CG1 VAL 695 61.120 26.268 66.113 1.00 68.63 0 ATOM 4750 CG2VAL 695 59.863 26.474 68.253 1.00 68.27 0 ATOM 4751 C VAL 695 63.24224.744 67.687 1.00 34.22 0 ATOM 4752 O VAL 695 64.070 25.548 68.123 1.0075.22 0 ATOM 4753 N PRO 696 63.599 23.730 66.873 1.00 2.00 0 ATOM 4754CD PRO 696 62.777 22.616 66.373 1.00 5.25 0 ATOM 4755 CA PRO 696 64.99823.549 66.462 1.00 2.00 0 ATOM 4756 CB PRO 696 64.997 22.163 65.803 1.005.25 0 ATOM 4757 CG PRO 696 63.768 21.486 66.381 1.00 5.25 0 ATOM 4758 CPRO 696 65.570 24.612 65.536 1.00 2.00 0 ATOM 4759 O PRO 696 64.87825.556 65.137 1.00 5.25 0 ATOM 4760 N ASP 697 66.850 24.431 65.214 1.0034.70 0 ATOM 4762 CA ASP 697 67.598 25.311 64.321 1.00 35.10 0 ATOM 4763CB ASP 697 69.098 24.951 64.376 1.00 81.36 0 ATOM 4764 CG ASP 697 69.63024.774 65.825 1.00 81.80 0 ATOM 4765 OD1 ASP 697 69.612 23.613 66.3471.00 0.89 0 ATOM 4766 OD2 ASP 697 70.075 25.792 66.436 1.00 0.05 0 ATOM4767 C ASP 697 67.037 25.099 62.894 1.00 36.63 0 ATOM 4768 O ASP 69767.014 26.022 62.069 1.00 0.75 0 ATOM 4769 N GLN 698 66.576 23.87762.620 1.00 8.48 0 ATOM 4771 CA GLN 698 65.997 23.510 61.335 1.00 2.00 0ATOM 4772 CB GLN 698 67.089 23.346 60.285 1.00 43.65 0 ATOM 4773 CG GLN698 68.191 22.376 60.649 1.00 44.93 0 ATOM 4774 CD GLN 698 69.158 22.16959.501 1.00 43.25 0 ATOM 4775 OE1 GLN 698 68.781 22.242 58.327 1.0045.90 0 ATOM 4776 NE2 GLN 698 70.411 21.911 59.830 1.00 44.14 0 ATOM4779 C GLN 698 65.211 22.216 61.482 1.00 2.32 0 ATOM 4780 O GLN 69865.396 21.486 62.452 1.00 42.62 0 ATOM 4781 N GLY 699 64.324 21.93960.530 1.00 2.00 0 ATOM 4783 CA GLY 699 63.510 20.728 60.576 1.00 2.00 0ATOM 4784 C GLY 699 62.046 21.059 60.346 1.00 2.00 0 ATOM 4785 O GLY 69961.726 22.204 60.013 1.00 2.00 0 ATOM 4786 N LEU 700 61.153 20.09060.545 1.00 2.00 0 ATOM 4788 CA LEU 700 59.700 20.297 60.352 1.00 2.00 0ATOM 4789 CB LEU 700 58.941 18.962 60.555 1.00 4.64 0 ATOM 4790 CG LEU700 57.436 18.836 60.273 1.00 8.84 0 ATOM 4791 CD1 LEU 700 57.219 18.88158.793 1.00 8.28 0 ATOM 4792 CD2 LEU 700 56.879 17.536 60.803 1.00 5.230 ATOM 4793 C LEU 700 59.086 21.400 61.247 1.00 2.00 0 ATOM 4794 O LEU700 58.365 22.261 60.763 1.00 8.77 0 ATOM 4795 N LEU 701 59.394 21.38162.540 1.00 12.05 0 ATOM 4797 CA LEU 701 58.860 22.362 63.477 1.00 11.210 ATOM 4798 CB LEU 701 59.278 22.039 64.908 1.00 2.00 0 ATOM 4799 CG LEU701 58.156 21.874 65.942 1.00 2.00 0 ATOM 4800 CD1 LEU 701 58.759 21.87967.354 1.00 2.00 0 ATOM 4801 CD2 LEU 701 57.124 22.994 65.800 1.00 2.000 ATOM 4802 C LEU 701 59.339 23.747 63.153 1.00 6.49 0 ATOM 4803 O LEU701 58.562 24.691 63.157 1.00 2.00 0 ATOM 4804 N CYS 702 60.626 23.88162.890 1.00 2.00 0 ATOM 4806 CA CYS 702 61.158 25.191 62.572 1.00 2.00 0ATOM 4807 CB CYS 702 62.646 25.117 62.269 1.00 10.82 0 ATOM 4608 SG CYS702 63.290 26.639 61.543 1.00 10.82 0 ATOM 4809 C CYS 702 60.442 25.78661.365 1.00 2.00 0 ATOM 4810 O CYS 702 60.022 26.954 61.384 1.00 10.82 0ATOM 4811 N ASP 703 60.292 24.980 60.316 1.00 2.00 0 ATOM 4813 CA ASP703 59.641 25.442 59.104 1.00 2.00 0 ATOM 4814 CB ASP 703 59.790 24.39158.010 1.00 9.22 0 ATOM 4815 CG ASP 703 61.251 24.076 57.698 1.00 9.98 0ATOM 4816 OD1 ASP 703 62.126 24.940 57.943 1.00 9.22 0 ATOM 4817 OD2 ASP703 61.537 22.956 57.213 1.00 9.22 0 ATOM 4818 C ASP 703 58.187 25.76059.377 1.00 2.00 0 ATOM 4819 O ASP 703 57.694 26.823 59.023 1.00 11.43 0ATOM 4820 N LEU 704 57.519 24.861 60.072 1.00 2.00 0 ATOM 4822 CA LEU704 56.113 25.051 60.391 1.00 2.00 0 ATOM 4823 CB LEU 704 55.630 23.94261.338 1.00 22.47 0 ATOM 4824 CG LEU 704 55.412 22.533 60.780 1.00 21.160 ATOM 4825 CD1 LEU 704 55.366 21.528 61.911 1.00 24.29 0 ATOM 4826 CD2LEU 704 54.132 22.489 59.973 1.00 22.40 0 ATOM 4827 C LEU 704 55.80926.417 61.006 1.00 2.00 0 ATOM 4828 O LEU 704 54.736 26.979 60.773 1.0015.38 0 ATOM 4829 N LEU 705 56.757 26.967 61.763 1.00 39.58 0 ATOM 4831CA LEU 705 56.538 28.248 62.439 1.00 39.58 0 ATOM 4832 CB LEU 705 56.88428.108 63.913 1.00 2.00 0 ATOM 4833 CG LEU 705 56.841 26.725 64.552 1.002.00 0 ATOM 4834 CD1 LEU 705 57.376 26.870 65.950 1.00 2.00 0 ATOM 4835CD2 LEU 705 55.446 26.156 64.566 1.00 2.00 0 ATOM 4836 C LEU 705 57.27929.466 61.900 1.00 39.58 0 ATOM 4837 O LEU 705 56.924 30.598 62.240 1.002.00 0 ATOM 4838 N TRP 706 58.307 29.237 61.086 1.00 2.00 0 ATOM 4840 CATRP 706 59.117 30.319 60.523 1.00 2.00 0 ATOM 4841 CB TRP 706 60.59430.025 60.777 1.00 23.95 0 ATOM 4842 CG TRP 706 61.025 30.261 62.1651.00 23.95 0 ATOM 4843 CD2 TRP 706 61.380 31.520 62.742 1.00 23.95 0ATOM 4844 CE2 TRP 706 61.735 31.278 64.083 1.00 23.95 0 ATOM 4845 CE3TRP 706 61.434 32.832 62.254 1.00 23.95 0 ATOM 4846 CD1 TRP 706 61.17329.329 63.150 1.00 23.95 0 ATOM 4847 NE1 TRP 706 61.599 29.932 64.3071.00 23.95 0 ATOM 4849 CZ2 TRP 706 62.141 32.303 64.946 1.00 23.95 0ATOM 4850 CZ3 TRP 706 61.835 33.850 63.108 1.00 23.95 0 ATOM 4851 CH2TRP 706 62.184 33.580 64.439 1.00 23.95 0 ATOM 4852 C TRP 706 58.94730.619 59.028 1.00 2.00 0 ATOM 4853 O TRP 706 59.186 31.751 58.598 1.0023.95 0 ATOM 4854 N SER 707 58.564 29.604 58.249 1.00 17.80 0 ATOM 4856CA SER 707 58.423 29.730 56.796 1.00 12.52 0 ATOM 4857 CB SER 707 58.03428.383 56.160 1.00 9.59 0 ATOM 4858 OG SER 707 56.693 28.010 56.444 1.008.78 0 ATOM 4860 C SER 707 57.459 30.806 56.328 1.00 18.86 0 ATOM 4861 OSER 707 56.521 31.179 57.035 1.00 6.18 0 ATOM 4862 N ASP 708 57.70031.303 55.124 1.00 7.67 0 ATOM 4864 CA ASP 708 56.860 32.328 54.547 1.007.67 0 ATOM 4865 CB ASP 708 57.546 33.676 54.656 1.00 8.53 0 ATOM 4866CG ASP 708 57.720 34.104 56.079 1.00 10.12 0 ATOM 4867 OD1 ASP 70858.844 34.049 56.597 1.00 10.18 0 ATOM 4868 OD2 ASP 708 56.717 34.48356.689 1.00 10.95 0 ATOM 4869 C ASP 708 56.609 31.998 53.104 1.00 7.67 0ATOM 4870 O ASP 708 57.461 31.432 52.444 1.00 11.43 0 ATOM 4871 N PRO709 55.425 32.328 52.596 1.00 2.00 0 ATOM 4872 OD PRO 709 54.346 33.00653.302 1.00 2.00 0 ATOM 4873 CA PRO 709 55.029 32.078 51.214 1.00 2.00 0ATOM 4874 CB PRO 709 53.507 32.036 51.283 1.00 2.00 0 ATOM 4875 CG PRO709 53.169 32.316 52.728 1.00 2.00 0 ATOM 4876 C PRO 709 55.475 33.26950.406 1.00 2.00 0 ATOM 4877 O PRO 709 55.071 34.393 50.712 1.00 2.00 0ATOM 4878 N ASP 710 56.306 33.046 49.393 1.00 2.57 0 ATOM 4880 CA ASP710 56.769 34.155 48.576 1.00 4.76 0 ATOM 4881 CB ASP 710 58.297 34.21548.537 1.00 18.70 0 ATOM 4882 CG ASP 710 58.824 35.631 48.303 1.00 26.430 ATOM 4883 OD1 ASP 710 58.070 36.497 47.793 1.00 26.58 0 ATOM 4884 OD2ASP 710 60.002 35.884 48.635 1.00 30.90 0 ATOM 4885 C ASP 710 56.21134.045 47.168 1.00 5.37 0 ATOM 4886 O ASP 710 56.454 33.056 46.467 1.0015.67 0 ATOM 4887 N LYS 711 55.449 35.074 46.789 1.00 10.56 0 ATOM 4889CA LYS 711 54.802 35.217 45.480 1.00 16.19 0 ATOM 4890 CB LYS 711 54.26636.661 45.368 1.00 35.75 0 ATOM 4891 CG LYS 711 53.743 37.109 44.0071.00 43.76 0 ATOM 4892 CD LYS 711 54.843 37.750 43.146 1.00 49.83 0 ATOM4893 CE LYS 711 55.459 38.976 43.819 1.00 54.70 0 ATOM 4894 NZ LYS 71156.632 39.514 43.064 1.00 58.93 0 ATOM 4898 C LYS 711 55.753 34.87744.331 1.00 16.03 0 ATOM 4899 O LYS 711 55.459 34.001 43.518 1.00 37.960 ATOM 4900 N ASP 712 56.894 35.560 44.282 1.00 2.00 0 ATOM 4902 CA ASP712 57.890 35.330 43.252 1.00 2.00 0 ATOM 4903 CB ASP 712 58.655 36.62942.927 1.00 75.12 0 ATOM 4904 CG ASP 712 59.362 37.231 44.134 1.00 75.410 ATOM 4905 OD1 ASP 712 58.723 38.013 44.866 1.00 77.74 0 ATOM 4906 OD2ASP 712 60.561 36.937 44.341 1.00 84.37 0 ATOM 4907 C ASP 712 58.86334.189 43.597 1.00 2.00 0 ATOM 4908 O ASP 712 60.083 34.366 43.607 1.0077.12 0 ATOM 4909 N VAL 713 58.298 33.016 43.866 1.00 27.09 0 ATOM 4911CA VAL 713 59.057 31.811 44.188 1.00 17.74 0 ATOM 4912 CB VAL 713 59.16631.579 45.727 1.00 2.00 0 ATOM 4913 CG1 VAL 713 59.124 30.098 46.0691.00 2.00 0 ATOM 4914 CG2 VAL 713 60.481 32.120 46.224 1.00 2.00 0 ATOM4915 C VAL 713 58.319 30.653 43.538 1.00 21.93 0 ATOM 4916 O VAL 71357.092 30.567 43.593 1.00 2.00 0 ATOM 4917 N LEU 714 59.063 29.76642.903 1.00 15.32 0 ATOM 4919 CA LEU 714 58.434 28.642 42.249 1.00 10.300 ATOM 4920 CB LEU 714 59.285 28.147 41.078 1.00 48.90 0 ATOM 4921 CGLEU 714 58.662 27.005 40.272 1.00 42.23 0 ATOM 4922 CD1 LEU 714 57.22927.357 39.909 1.00 43.72 0 ATOM 4923 CD2 LEU 714 59.478 26.753 39.0251.00 42.86 0 ATOM 4924 C LEU 714 58.224 27.530 43.235 1.00 13.23 0 ATOM4925 O LEU 714 57.096 27.099 43.445 1.00 44.72 0 ATOM 4926 N GLY 71559.322 27.075 43.834 1.00 76.36 0 ATOM 4928 CA GLY 715 59.263 25.99544.800 1.00 76.36 0 ATOM 4929 C GLY 715 59.630 26.420 46.206 1.00 76.360 ATOM 4930 O GLY 715 58.814 26.974 46.929 1.00 19.62 0 ATOM 4931 N TRP716 60.875 26.183 46.583 1.00 4.87 0 ATOM 4932 CA TRP 716 61.365 26.50347.918 1.00 4.87 0 ATOM 4934 CB TRP 716 61.944 25.241 48.554 1.00 2.00 0ATOM 4935 CG TRP 716 60.884 24.317 48.932 1.00 2.00 0 ATOM 4936 CD2 TRP716 59.936 24.529 49.962 1.00 2.00 0 ATOM 4937 CE2 TRP 716 59.050 23.43949.943 1.00 2.00 0 ATOM 4938 CE3 TRP 716 59.750 25.541 50.909 1.00 2.000 ATOM 4939 CD1 TRP 716 60.562 23.134 48.343 1.00 2.00 0 ATOM 4940 NE1TRP 716 59.454 22.597 48.940 1.00 2.00 0 ATOM 4942 CZ2 TRP 716 57.99423.334 50.830 1.00 2.00 0 ATOM 4943 CZ3 TRP 716 58.715 25.438 51.7831.00 2.00 0 ATOM 4944 CH2 TRP 716 57.843 24.343 51.743 1.00 2.00 0 ATOM4945 C TRP 716 62.406 27.594 47.954 1.00 4.87 0 ATOM 4946 O TRP 71663.596 27.315 47.871 1.00 2.00 0 ATOM 4947 N GLY 717 61.968 28.83748.067 1.00 2.00 0 ATOM 4949 CA GLY 717 62.911 29.937 48.124 1.00 2.00 0ATOM 4950 C GLY 717 63.725 30.031 49.414 1.00 2.00 0 ATOM 4951 O GLY 71763.443 29.343 50.404 1.00 2.00 0 ATOM 4952 N GLU 718 64.752 30.88049.387 1.00 4.99 0 ATOM 4954 CA GLU 718 65.606 31.113 50.538 1.00 8.49 0ATOM 4955 CB GLU 718 66.980 31.619 50.092 1.00 86.14 0 ATOM 4956 CG GLU718 68.026 31.716 51.211 1.00 89.35 0 ATOM 4957 CD GLU 718 68.627 30.37151.633 1.00 87.67 0 ATOM 4958 OE1 GLU 718 69.392 30.358 52.624 1.0095.59 0 ATOM 4959 OE2 GLU 718 68.353 29.336 50.986 1.00 88.92 0 ATOM4960 C GLU 718 64.883 32.182 51.340 1.00 9.60 0 ATOM 4961 O GLU 71864.597 33.269 50.828 1.00 90.23 0 ATOM 4962 N ASN 719 64.560 31.86352.590 1.00 32.81 0 ATOM 4964 CA ASN 719 63.848 32.804 53.444 1.00 33.930 ATOM 4965 CB ASN 719 63.159 32.083 54.591 1.00 14.30 0 ATOM 4966 CGASN 719 62.159 32.965 55.296 1.00 14.30 0 ATOM 4967 OD1 ASN 719 62.53333.912 55.983 1.00 14.30 0 ATOM 4968 ND2 ASN 719 60.879 32.676 55.1101.00 14.30 0 ATOM 4971 C ASN 719 64.781 33.855 54.007 1.00 33.45 0 ATOM4972 O ASN 719 65.825 33.529 54.566 1.00 14.30 0 ATOM 4973 N ASP 72064.388 35.117 53.878 1.00 42.61 0 ATOM 4975 CA ASP 720 65.212 36.22254.351 1.00 47.00 0 ATOM 4976 CB ASP 720 64.744 37.535 53.706 1.00 77.300 ATOM 4977 CG ASP 720 65.025 37.574 52.200 1.00 92.85 0 ATOM 4978 OD1ASP 720 66.104 38.071 51.801 1.00 92.39 0 ATOM 4979 OD2 ASP 720 64.17237.094 51.416 1.00 90.51 0 ATOM 4980 C ASP 720 65.328 36.352 55.869 1.0039.89 0 ATOM 4981 O ASP 720 66.255 36.986 56.370 1.00 74.64 0 ATOM 4992N ARG 721 64.411 35.728 56.602 1.00 13.83 0 ATOM 4994 CA ARG 721 64.44635.759 58.060 1.00 12.18 0 ATOM 4985 CB ARG 721 63.262 35.007 58.6491.00 15.37 0 ATOM 4986 CG ARG 721 61.946 35.693 58.572 1.00 7.28 0 ATOM4987 CD ARG 721 60.950 34.836 59.300 1.00 7.28 0 ATOM 4988 NE ARG 72159.593 35.306 59.103 1.00 8.88 0 ATOM 4990 CZ ARG 721 59.051 36.31559.766 1.00 9.83 0 ATOM 4991 NH1 ARG 721 59.751 36.965 60.690 1.00 8.950 ATOM 4994 NH2 ARG 721 57.809 36.682 59.485 1.00 8.62 0 ATOM 4997 C ARG721 65.710 35.093 58.592 1.00 8.44 0 ATOM 4998 O ARG 721 65.982 35.14259.798 1.00 10.25 0 ATOM 4999 N GLY 722 66.449 34.437 57.697 1.00 4.85 0ATOM 5001 CA GLY 722 67.668 33.752 58.083 1.00 4.85 0 ATOM 5002 C GLY722 67.392 32.344 58.592 1.00 4.85 0 ATOM 5003 O GLY 722 68.305 31.65559.052 1.00 76.68 0 ATOM 5004 N VAL 723 66.134 31.918 58.508 1.00 10.850 ATOM 5006 CA VAL 723 65.717 30.591 58.958 1.00 10.85 0 ATOM 5007 CBVAL 723 65.259 30.574 60.471 1.00 2.00 0 ATOM 5008 CG1 VAL 723 66.44930.400 61.386 1.00 2.00 0 ATOM 5009 CG2 VAL 723 64.504 31.858 60.8251.00 2.00 0 ATOM 5010 C VAL 723 64.546 30.100 58.105 1.00 10.85 0 ATOM5011 O VAL 723 63.667 30.883 57.714 1.00 2.00 0 ATOM 5012 N SER 72464.541 28.802 57.824 1.00 42.15 0 ATOM 5014 CA SER 724 63.479 28.19357.040 1.00 42.15 0 ATOM 5015 CB SER 724 62.127 28.609 57.623 1.00 2.000 ATOM 5016 OG SER 724 61.077 27.808 57.106 1.00 2.00 0 ATOM 5018 C SER724 63.583 28.588 55.561 1.00 42.15 0 ATOM 5019 O SER 724 64.680 28.82055.049 1.00 2.00 0 ATOM 5020 N PHE 725 62.454 28.643 54.863 1.00 2.00 0ATOM 5022 CA PHE 725 62.464 29.007 53.457 1.00 2.00 0 ATOM 5023 CB PHE725 62.461 27.779 52.541 1.00 2.00 0 ATOM 5024 CG PHE 725 62.891 26.55153.205 1.00 2.00 0 ATOM 5025 CD1 PHE 725 62.047 25.916 54.089 1.00 2.000 ATOM 5026 CD2 PHE 725 64.151 26.044 52.984 1.00 2.00 0 ATOM 5027 CE1PHE 725 62.458 24.791 54.750 1.00 2.00 0 ATOM 5028 CE2 PHE 725 64.57824.906 53.646 1.00 2.00 0 ATOM 5029 CZ PHE 725 63.733 24.280 54.530 1.002.00 0 ATOM 5030 C PHE 725 61.222 29.787 53.146 1.00 2.00 0 ATOM 5031 OPHE 725 60.382 30.040 54.009 1.00 2.00 0 ATOM 5032 N THR 726 61.13230.162 51.886 1.00 34.54 0 ATOM 5034 CA THR 726 60.009 30.877 51.3571.00 30.82 0 ATOM 5035 CB THR 726 60.468 32.154 50.661 1.00 2.00 0 ATOM5036 OG1 THR 726 61.880 32.098 50.406 1.00 2.00 0 ATOM 5038 CG2 THR 72660.222 33.327 51.561 1.00 2.00 0 ATOM 5039 C THR 726 59.450 29.86650.382 1.00 34.20 0 ATOM 5040 O THR 726 60.201 29.208 49.684 1.00 2.00 0ATOM 5041 N PHE 727 58.144 29.685 50.375 1.00 2.00 0 ATOM 5043 CA PHE727 57.555 28.712 49.483 1.00 2.00 0 ATOM 5044 CB PHE 727 56.852 27.61250.275 1.00 9.97 0 ATOM 5045 CG PHE 727 55.698 28.096 51.105 1.00 15.330 ATOM 5046 CD1 PHE 727 54.394 27.922 50.666 1.00 9.89 0 ATOM 5047 CD2PHE 727 55.916 28.726 52.323 1.00 15.65 0 ATOM 5048 CE1 PHE 727 53.33828.360 51.418 1.00 11.89 0 ATOM 5049 CE2 PHE 727 54.852 29.171 53.0871.00 6.87 0 ATOM 5050 CZ PHE 727 53.563 28.986 52.631 1.00 11.72 0 ATOM5051 C PHE 727 56.580 29.398 48.553 1.00 2.00 0 ATOM 5052 O PHE 72755.848 30.312 48.982 1.00 18.77 0 ATOM 5053 N GLY 728 56.576 28.95647.289 1.00 13.14 0 ATOM 5055 CA GLY 728 55.709 29.542 46.277 1.00 12.120 ATOM 5056 C GLY 728 54.348 28.894 46.174 1.00 13.69 0 ATOM 5057 O GLY728 54.062 27.913 46.862 1.00 2.00 0 ATOM 5058 N ALA 729 53.513 29.43645.292 1.00 33.39 0 ATOM 5060 CA ALA 729 52.161 28.916 45.080 1.00 33.110 ATOM 5061 CB ALA 729 51.375 29.862 44.200 1.00 16.31 0 ATOM 5062 C ALA729 52.139 27.507 44.485 1.00 31.34 0 ATOM 5063 O ALA 729 51.143 26.79644.600 1.00 16.31 0 ATOM 5064 N GLU 730 53.221 27.105 43.831 1.00 22.530 ATOM 5066 CA GLU 730 53.284 25.761 43.281 1.00 26.87 0 ATOM 5067 CBGLU 730 54.622 25.551 42.570 1.00 59.75 0 ATOM 5068 CG GLU 730 54.89324.117 42.142 1.00 62.63 0 ATOM 5069 CD GLU 730 56.138 23.984 41.2831.00 68.00 0 ATOM 5070 OE1 GLU 730 57.213 23.624 41.822 1.00 74.97 0ATOM 5071 OE2 GLU 730 56.034 24.238 40.063 1.00 67.31 0 ATOM 5072 C GLU730 53.140 24.781 44.446 1.00 24.85 0 ATOM 5073 O GLU 730 52.285 23.89944.425 1.00 58.27 0 ATOM 5074 N VAL 731 53.958 24.989 45.477 1.00 24.130 ATOM 5076 CA VAL 731 53.985 24.157 46.679 1.00 19.42 0 ATOM 5077 CBVAL 731 55.079 24.628 47.645 1.00 19.11 0 ATOM 5078 CG1 VAL 731 55.15923.699 48.824 1.00 19.11 0 ATOM 5079 CG2 VAL 731 56.412 24.691 46.9421.00 19.11 0 ATOM 5080 C VAL 731 52.659 24.165 47.423 1.00 16.89 0 ATOM5081 O VAL 731 52.210 23.128 47.900 1.00 19.11 0 ATOM 5082 N VAL 73252.035 25.332 47.525 1.00 15.54 0 ATOM 5084 CA VAL 732 50.750 25.44948.209 1.00 15.54 0 ATOM 5085 CB VAL 732 50.254 26.898 48.240 1.00 20.170 ATOM 5086 CG1 VAL 732 48.907 26.985 48.962 1.00 20.17 0 ATOM 5087 CG2VAL 732 51.272 27.764 48.902 1.00 20.17 0 ATOM 5088 C VAL 732 49.65324.609 47.554 1.00 15.54 0 ATOM 5089 O VAL 732 49.011 23.791 48.222 1.0020.17 0 ATOM 5090 N ALA 733 49.437 24.822 46.253 1.00 16.29 0 ATOM 5092CA ALA 733 48.408 24.109 45.501 1.00 16.29 0 ATOM 5093 CB ALA 733 48.26024.717 44.131 1.00 17.33 0 ATOM 5094 C ALA 733 48.703 22.618 45.390 1.0016.29 0 ATOM 5095 O ALA 733 47.776 21.795 45.349 1.00 17.78 0 ATOM 5096N LYS 734 49.996 22.287 45.348 1.00 2.00 0 ATOM 5098 CA LYS 734 50.49920.905 45.259 1.00 2.00 0 ATOM 5099 CB LYS 734 52.012 20.936 45.015 1.0023.03 0 ATOM 5100 CG LYS 734 52.507 20.244 43.759 1.00 25.55 0 ATOM 5101CD LYS 734 52.696 21.212 42.594 1.00 35.13 0 ATOM 5102 CE LYS 734 53.61320.608 41.521 1.00 40.96 0 ATOM 5103 NZ LYS 734 55.006 20.311 42.0011.00 46.09 0 ATOM 5107 C LYS 734 50.222 20.146 46.578 1.00 2.00 0 ATOM5108 O LYS 734 49.995 18.926 46.591 1.00 16.68 0 ATOM 5109 N PHE 73550.263 20.915 47.670 1.00 36.96 0 ATOM 5111 CA PHE 735 50.036 20.47849.047 1.00 33.73 0 ATOM 5112 CB PHE 735 50.606 21.554 49.991 1.00 8.600 ATOM 5113 CG PHE 735 50.320 21.323 51.465 1.00 8.60 0 ATOM 5114 CD1PHE 735 50.955 20.298 52.169 1.00 8.60 0 ATOM 5115 CD2 PHE 735 49.41222.135 52.142 1.00 8.60 0 ATOM 5116 CE1 PHE 735 50.690 20.086 53.5121.00 8.60 0 ATOM 5117 CE2 PHE 735 49.143 21.929 53.483 1.00 8.60 0 ATOM5118 CZ PHE 735 49.784 20.900 54.168 1.00 8.60 0 ATOM 5119 C PHE 73546.546 20.256 49.334 1.00 34.15 0 ATOM 5120 O PHE 735 48.151 19.18149.801 1.00 8.60 0 ATOM 5121 N LEU 736 47.731 21.281 49.075 1.00 2.00 0ATOM 5123 CA LEU 736 46.289 21.208 49.299 1.00 2.00 0 ATOM 5124 CB LEU736 45.599 22.451 48.770 1.00 2.00 0 ATOM 5125 CG LEU 736 45.937 23.76049.456 1.00 2.00 0 ATOM 5126 CD1 LEU 736 45.314 24.887 48.689 1.00 2.000 ATOM 5127 CD2 LEU 736 45.415 23.743 50.896 1.00 2.00 0 ATOM 5128 C LEU736 45.741 20.014 48.567 1.00 2.00 0 ATOM 5129 O LEU 736 44.986 19.21549.121 1.00 2.00 0 ATOM 5130 N HIS 737 46.147 19.906 47.308 1.00 25.73 0ATOM 5132 CA HIS 737 45.747 18.818 46.427 1.00 25.73 0 ATOM 5133 CB HIS737 46.423 19.037 45.057 1.00 69.97 0 ATOM 5134 CG HIS 737 46.638 17.78444.263 1.00 68.65 0 ATOM 5135 CD2 HIS 737 47.779 17.169 43.868 1.0071.13 0 ATOM 5136 ND1 HIS 737 45.601 17.012 43.783 1.00 76.45 0 ATOM5138 CE1 HIS 737 46.093 15.974 43.129 1.00 76.42 0 ATOM 5139 NE2 HIS 73747.412 16.046 43.166 1.00 74.54 0 ATOM 5141 C HIS 737 46.088 17.44047.037 1.00 25.73 0 ATOM 5142 O HIS 737 45.223 16.570 47.166 1.00 61.280 ATOM 5143 N LYS 738 47.347 17.266 47.422 1.00 11.82 0 ATOM 5145 CA LYS738 47.836 16.028 48.010 1.00 10.83 0 ATOM 5146 CB LYS 738 49.343 16.16348.256 1.00 10.15 0 ATOM 5147 CG LYS 738 49.999 15.111 49.119 1.00 13.960 ATOM 5148 CD LYS 738 51.516 15.253 49.005 1.00 16.98 0 ATOM 5149 CELYS 738 52.278 14.809 50.274 1.00 14.10 0 ATOM 5150 NZ LYS 738 52.15413.362 50.650 1.00 13.23 0 ATOM 5154 C LYS 738 47.121 15.655 49.301 1.0017.32 0 ATOM 5155 O LYS 738 47.038 14.473 49.640 1.00 16.32 0 ATOM 5156N HIS 739 46.598 16.642 50.028 1.00 2.00 0 ATOM 5158 CA HIS 739 45.93516.319 51.284 1.00 2.00 0 ATOM 5159 CB HIS 739 46.689 16.965 52.460 1.009.90 0 ATOM 5160 CG HIS 739 48.099 16.482 52.597 1.00 8.91 0 ATOM 5161CD2 HIS 739 48.603 15.318 53.077 1.00 3.85 0 ATOM 5162 ND1 HIS 73949.182 17.212 52.148 1.00 7.65 0 ATOM 5164 CE1 HIS 739 50.291 16.51752.340 1.00 6.50 0 ATOM 5165 NE2 HIS 739 49.966 15.364 52.901 1.00 3.850 ATOM 5167 C HIS 739 44.442 16.598 51.378 1.00 2.00 0 ATOM 5168 O HIS739 43.913 16.722 52.477 1.00 18.58 0 ATOM 5169 N ASP 740 43.763 16.67850.236 1.00 2.00 0 ATOM 5171 CA ASP 740 42.314 16.912 50.191 1.00 2.00 0ATOM 5172 CB ASP 740 41.567 15.695 50.758 1.00 37.08 0 ATOM 5173 CG ASP740 42.092 14.367 50.206 1.00 45.58 0 ATOM 5174 OD1 ASP 740 42.97413.754 50.856 1.00 45.42 0 ATOM 5175 OD2 ASP 740 41.622 13.932 49.1291.00 41.93 0 ATOM 5176 C ASP 740 41.900 18.170 50.955 1.00 2.00 0 ATOM5177 O ASP 740 40.773 18.286 51.432 1.00 36.93 0 ATOM 5178 N LEU 74142.824 19.113 51.045 1.00 2.00 0 ATOM 5180 CA LEU 741 42.610 20.35751.764 1.00 2.00 0 ATOM 5181 CB LEU 741 43.920 20.818 52.415 1.00 2.00 0ATOM 5182 CG LEU 741 44.572 19.861 53.388 1.00 2.00 0 ATOM 5183 CD1 LEU741 45.906 20.394 53.859 1.00 2.00 0 ATOM 5184 CD2 LEU 741 43.620 19.67354.519 1.00 2.00 0 ATOM 5185 C LEU 741 42.122 21.433 50.818 1.00 2.00 0ATOM 5186 O LEU 741 42.261 21.301 49.611 1.00 2.00 0 ATOM 5187 N ASP 74241.588 22.510 51.385 1.00 2.00 0 ATOM 5189 CA ASP 742 41.080 23.62050.607 1.00 2.00 0 ATOM 5190 CB ASP 742 39.605 23.858 50.909 1.00 21.530 ATOM 5191 CG ASP 742 38.717 22.741 50.433 1.00 26.90 0 ATOM 5192 OD1ASP 742 39.159 21.898 49.622 1.00 27.19 0 ATOM 5193 OD2 ASP 742 37.55522.714 50.876 1.00 25.88 0 ATOM 5194 C ASP 742 41.810 24.913 50.887 1.002.00 0 ATOM 5195 O ASP 742 42.064 25.698 49.970 1.00 14.77 0 ATOM 5196 NLEU 743 42.141 25.152 52.149 1.00 5.78 0 ATOM 5198 CA LEU 743 42.78526.407 52.509 1.00 5.78 0 ATOM 5199 CB LEU 743 41.744 27.312 53.185 1.002.00 0 ATOM 5200 CG LEU 743 41.689 28.847 53.088 1.00 2.00 0 ATOM 5201CD1 LEU 743 40.947 29.364 54.297 1.00 2.00 0 ATOM 5202 CD2 LEU 74343.051 29.469 53.053 1.00 2.00 0 ATOM 5203 C LEU 743 41.919 26.15253.483 1.00 5.78 0 ATOM 5204 O LEU 743 43.973 25.091 54.088 1.00 2.00 0ATOM 5205 N ILE 744 44.837 27.104 53.599 1.00 2.00 0 ATOM 5207 CA ILE744 45.916 27.031 54.574 1.00 2.00 0 ATOM 5208 CB ILE 744 47.338 27.04353.968 1.00 2.00 0 ATOM 5209 CG2 ILE 744 48.360 27.214 55.089 1.00 2.000 ATOM 5210 CG1 ILE 744 47.637 25.754 53.204 1.00 2.00 0 ATOM 5211 CD1ILE 744 49.117 25.602 52.856 1.00 2.00 0 ATOM 5212 C ILE 744 45.77028.330 55.356 1.00 2.00 0 ATOM 5213 O ILE 744 45.830 29.423 54.788 1.002.00 0 ATOM 5214 N CYS 745 45.557 28.226 56.655 1.00 71.57 0 ATOM 5216CA CYS 745 45.426 29.418 57.462 1.00 66.70 0 ATOM 5217 CB CYS 745 44.20429.301 58.363 1.00 17.55 0 ATOM 5218 SG CYS 745 43.454 30.878 58.7031.00 23.65 0 ATOM 5219 C CYS 745 46.700 29.572 58.281 1.00 69.12 0 ATOM5220 O CYS 745 47.093 28.667 59.017 1.00 21.37 0 ATOM 5221 N ARG 74647.363 30.706 58.121 1.00 2.00 0 ATOM 5223 CA ARG 746 48.594 30.98258.837 1.00 2.00 0 ATOM 5224 CB ARG 746 49.810 30.672 57.944 1.00 2.00 0ATOM 5225 CG ARG 746 49.860 31.394 56.609 1.00 2.00 0 ATOM 5226 CD ARG746 50.792 32.619 56.617 1.00 2.00 0 ATOM 5227 NE ARG 746 52.220 32.29356.560 1.00 2.00 0 ATOM 5229 CZ ARG 746 53.212 33.184 56.626 1.00 2.00 0ATOM 5230 NH1 ARG 746 52.956 34.479 56.738 1.00 2.00 0 ATOM 5233 NH2 ARG746 54.468 32.778 56.603 1.00 2.00 0 ATOM 5236 C ARG 746 48.597 32.43959.271 1.00 2.00 0 ATOM 5237 O ARG 746 47.739 33.220 58.847 1.00 2.00 0ATOM 5238 N ALA 747 49.524 32.802 60.146 1.00 2.00 0 ATOM 5240 CA ALA747 49.635 34.185 60.595 1.00 2.00 0 ATOM 5241 CB ALA 747 49.410 34.26462.106 1.00 2.00 0 ATOM 5242 C ALA 747 51.073 34.588 60.193 1.00 2.00 0ATOM 5243 O ALA 747 51.378 34.658 58.999 1.00 2.00 0 ATOM 5244 N HIS 74851.943 34.854 61.168 1.00 2.00 0 ATOM 5246 CA HIS 748 53.359 35.16860.939 1.00 2.00 0 ATOM 5247 C HIS 748 53.812 36.462 60.309 1.00 2.00 0ATOM 5248 O HIS 748 54.820 37.004 60.744 1.00 2.00 0 ATOM 5249 CB HIS748 54.032 34.028 60.187 1.00 2.00 0 ATOM 5250 CG HIS 748 55.503 33.91660.429 1.00 2.00 0 ATOM 5251 ND1 HIS 748 56.023 33.803 61.694 1.00 2.000 ATOM 5252 CE1 HIS 748 57.308 33.548 61.526 1.00 2.00 0 ATOM 5253 CD2HIS 748 56.493 33.737 59.527 1.00 2.00 0 ATOM 5254 NE2 HIS 748 57.63433.500 60.237 1.00 2.00 0 ATOM 5256 N GLN 749 53.116 36.962 59.302 1.002.00 0 ATOM 5258 CA GLN 749 53.556 38.197 58.677 1.00 2.00 0 ATOM 5259CB GLN 749 53.964 37.940 57.249 1.00 27.13 0 ATOM 5260 CG GLN 749 55.25737.222 57.143 1.00 28.03 0 ATOM 5261 CD GLN 749 55.586 36.915 55.7201.00 30.37 0 ATOM 5262 OE1 GLN 749 56.425 37.580 55.107 1.00 30.92 0ATOM 5263 NE2 GLN 749 54.928 35.898 55.173 1.00 30.08 0 ATOM 5266 C GLN749 52.556 39.310 58.708 1.00 2.00 0 ATOM 5267 O GLN 749 51.388 39.10658.407 1.00 25.39 0 ATOM 5268 N VAL 750 53.038 40.498 59.069 1.00 6.34 0ATOM 5270 CA VAL 750 52.193 41.682 59.157 1.00 6.34 0 ATOM 5271 CB VAL750 52.968 42.896 59.764 1.00 12.20 0 ATOM 5272 CG1 VAL 750 54.18343.216 58.935 1.00 12.20 0 ATOM 5273 CG2 VAL 750 52.055 44.109 59.8721.00 12.20 0 ATOM 5274 C VAL 750 51.672 42.018 57.771 1.00 6.34 0 ATOM5275 O VAL 750 52.471 42.219 56.850 1.00 12.20 0 ATOM 5276 N VAL 75150.343 42.001 57.617 1.00 25.10 0 ATOM 5278 CA VAL 751 49.685 42.32756.348 1.00 25.10 0 ATOM 5279 CB VAL 751 48.617 41.298 55.902 1.00 2.000 ATOM 5280 CG1 VAL 751 49.271 39.971 55.524 1.00 2.00 0 ATOM 5281 CG2VAL 751 47.570 41.138 56.976 1.00 2.00 0 ATOM 5282 C VAL 751 48.99643.654 56.540 1.00 25.10 0 ATOM 5283 O VAL 751 48.606 43.995 57.646 1.002.00 0 ATOM 5284 N GLU 752 48.820 44.389 55.453 1.00 2.00 0 ATOM 5286 CAGLU 752 48.219 45.709 55.526 1.00 2.00 0 ATOM 5287 CB GLU 752 48.43046.435 54.190 1.00 39.24 0 ATOM 5288 CG GLU 752 49.867 46.349 53.6281.00 75.32 0 ATOM 5289 CD GLU 752 50.962 46.804 54.611 1.00 75.31 0 ATOM5290 OE1 GLU 752 52.007 46.121 54.687 1.00 74.90 0 ATOM 5291 OE2 GLU 75250.792 47.837 55.300 1.00 91.76 0 ATOM 5292 C GLU 752 46.744 45.71655.913 1.00 2.00 0 ATOM 5293 O GLU 752 46.326 46.469 56.796 1.00 38.94 0ATOM 5294 N ASP 753 45.968 44.860 55.259 1.00 31.76 0 ATOM 5296 CA ASP753 44.527 44.776 55.491 1.00 34.73 0 ATOM 5297 CB ASP 753 43.787 44.47054.170 1.00 79.26 0 ATOM 5298 CG ASP 753 44.539 43.487 53.251 1.00 85.100 ATOM 5299 OD1 ASP 753 44.040 43.240 52.130 1.00 88.01 0 ATOM 5300 OD2ASP 753 45.613 42.960 53.619 1.00 92.40 0 ATOM 5301 C ASP 753 44.02843.853 56.606 1.00 31.43 0 ATOM 5302 O ASP 753 42.824 43.759 56.835 1.0064.64 0 ATOM 5303 N GLY 754 44.940 43.188 57.309 1.00 8.60 0 ATOM 5305CA GLY 754 44.535 42.309 58.393 1.00 5.10 0 ATOM 5306 C GLY 754 44.57740.849 58.007 1.00 3.66 0 ATOM 5307 O GLY 754 44.781 39.959 58.853 1.002.00 0 ATOM 5308 N TYR 755 44.361 40.611 56.718 1.00 27.09 0 ATOM 5310CA TYR 755 44.388 39.282 56.129 1.00 27.09 0 ATOM 5311 CB TYR 755 43.00438.632 56.128 1.00 28.14 0 ATOM 5312 CG TYR 755 41.976 39.338 55.2791.00 25.47 0 ATOM 5313 CD1 TYR 755 41.236 40.393 55.797 1.00 23.65 0ATOM 5314 CE1 TYR 755 40.276 41.049 55.034 1.00 28.13 0 ATOM 5315 CD2TYR 755 41.736 38.949 53.964 1.00 30.24 0 ATOM 5316 CE2 TYR 755 40.77439.601 53.187 1.00 27.11 0 ATOM 5317 CZ TYR 755 40.046 40.653 53.7321.00 32.27 0 ATOM 5318 OH TYR 755 39.086 41.311 52.996 1.00 28.90 0 ATOM5320 C TYR 755 44.844 39.530 54.713 1.00 27.09 0 ATOM 5321 O TYR 75544.772 40.654 54.238 1.00 25.53 0 ATOM 5322 N GLU 756 45.288 38.48354.035 1.00 8.71 0 ATOM 5324 CA GLU 756 45.787 38.617 52.676 1.00 8.71 0ATOM 5325 CB GLU 756 47.170 39.251 52.740 1.00 5.76 0 ATOM 5326 CG GLU756 47.832 39.557 51.436 1.00 16.72 0 ATOM 5327 CD GLU 756 49.127 40.30351.671 1.00 23.34 0 ATOM 5328 OE1 GLU 756 49.074 41.554 51.801 1.0025.72 0 ATOM 5329 OE2 GLU 756 50.194 39.636 51.751 1.00 25.15 0 ATOM5330 C GLU 756 45.853 37.240 52.044 1.00 8.71 0 ATOM 5331 O GLU 75646.400 36.306 52.638 1.00 6.14 0 ATOM 5332 N PHE 757 45.269 37.10550.858 1.00 28.33 0 ATOM 5334 CA PHE 757 45.270 35.822 50.174 1.00 28.330 ATOM 5335 CB PHE 757 44.055 35.683 49.253 1.00 2.00 0 ATOM 5336 CG PHE757 42.748 35.612 49.982 1.00 2.00 0 ATOM 5337 CD1 PHE 757 41.961 36.74750.137 1.00 2.00 0 ATOM 5338 CD2 PHE 757 42.306 34.407 50.535 1.00 2.000 ATOM 5339 CE1 PHE 757 40.751 36.695 50.833 1.00 2.00 0 ATOM 5340 CE2PHE 757 41.092 34.337 51.238 1.00 2.00 0 ATOM 5341 CZ PHE 757 40.31535.489 51.385 1.00 2.00 0 ATOM 5342 C PHE 757 46.547 35.639 49.387 1.0028.33 0 ATOM 5343 O PHE 757 47.220 36.602 49.033 1.00 2.00 0 ATOM 5344 NPHE 758 46.893 34.387 49.150 1.00 2.00 0 ATOM 5346 CA PHE 758 48.07534.050 48.397 1.00 2.00 0 ATOM 5347 CB PHE 758 49.241 33.789 49.317 1.0010.52 0 ATOM 5348 CG PHE 758 50.450 33.263 48.618 1.00 7.54 0 ATOM 5349CD1 PHE 758 51.404 34.131 48.100 1.00 7.91 0 ATOM 5350 CD2 PHE 75850.654 31.895 48.492 1.00 7.80 0 ATOM 5351 CE1 PHE 758 52.546 33.64247.467 1.00 11.14 0 ATOM 5352 CE2 PHE 758 51.796 31.398 47.858 1.0011.57 0 ATOM 5353 CZ PHE 758 52.741 32.272 47.347 1.00 10.83 0 ATOM 5354C PHE 758 47.725 32.786 47.636 1.00 2.00 0 ATOM 5355 O PHE 758 46.82632.034 48.042 1.00 13.46 0 ATOM 5356 N ALA 759 48.415 32.573 46.518 1.0017.88 0 ATOM 5358 CA ALA 759 48.196 31.407 45.573 1.00 17.63 0 ATOM 5359CB ALA 759 48.767 30.155 46.334 1.00 2.00 0 ATOM 5360 C ALA 759 46.73331.180 45.314 1.00 19.54 0 ATOM 5361 O ALA 759 46.187 30.106 45.547 1.002.00 0 ATOM 5362 N LYS 760 46.086 32.205 44.786 1.00 3.38 0 ATOM 5364 CALYS 760 44.699 32.076 44.341 1.00 3.38 0 ATOM 5365 CB LYS 760 44.63931.068 43.184 1.00 64.10 0 ATOM 5366 CG LYS 760 45.654 31.316 42.0621.00 64.10 0 ATOM 5367 CD LYS 760 45.843 30.068 41.205 1.00 64.10 0 ATOM5368 CE LYS 760 44.506 29.540 40.699 1.00 64.10 0 ATOM 5369 NZ LYS 76044.621 28.293 39.896 1.00 64.10 0 ATOM 5373 C LYS 760 43.715 31.65945.432 1.00 3.38 0 ATOM 5374 O LYS 760 42.953 30.702 45.256 1.00 64.10 0ATOM 5375 N ARG 761 43.751 32.376 46.557 1.00 16.83 0 ATOM 5377 CA ARG761 42.869 32.139 47.721 1.00 16.83 0 ATOM 5378 CB ARG 761 41.399 32.15447.274 1.00 38.86 0 ATOM 5379 CG ARG 761 41.012 33.375 46.458 1.00 38.860 ATOM 5380 CD ARG 761 40.550 34.493 47.334 1.00 38.86 0 ATOM 5381 NEARG 761 40.635 35.785 46.669 1.00 38.86 0 ATOM 5383 CZ ARG 761 39.79936.793 46.898 1.00 38.86 0 ATOM 5384 NH1 ARG 761 38.798 36.642 47.7671.00 38.86 0 ATOM 5387 NH2 ARG 761 39.988 37.963 46.291 1.00 38.86 0ATOM 5390 C ARG 761 43.145 30.844 48.508 1.00 16.83 0 ATOM 5391 O ARG761 42.596 30.639 49.585 1.00 38.86 0 ATOM 5392 N GLN 762 44.021 30.00247.971 1.00 24.54 0 ATOM 5394 CA GLN 762 44.359 28.718 48.559 1.00 24.540 ATOM 5395 CB GLN 762 45.167 27.908 47.553 1.00 19.81 0 ATOM 5396 CGGLN 762 44.502 27.794 46.173 1.00 19.81 0 ATOM 5397 CD GLN 762 45.30926.959 45.196 1.00 19.81 0 ATOM 5398 OE1 GLN 762 45.111 25.745 45.0971.00 19.81 0 ATOM 5399 NE2 GLN 762 46.230 27.596 44.476 1.00 19.81 0ATOM 5402 C GLN 762 45.126 28.856 49.857 1.00 24.54 0 ATOM 5403 O GLN762 45.278 27.893 50.599 1.00 19.81 0 ATOM 5404 N LEU 763 45.631 30.05350.119 1.00 2.00 0 ATOM 5406 CA LEU 763 46.354 30.326 51.353 1.00 2.00 0ATOM 5407 CB LEU 763 47.882 30.319 51.127 1.00 13.68 0 ATOM 5408 CG LEU763 48.847 30.427 52.335 1.00 13.68 0 ATOM 5409 CD1 LEU 763 50.11729.641 52.066 1.00 13.68 0 ATOM 5410 CD2 LEU 763 49.200 31.871 52.6331.00 13.68 0 ATOM 5411 C LEU 763 45.893 31.705 51.809 1.00 2.00 0 ATOM5412 O LEU 763 45.654 32.595 50.981 1.00 13.68 0 ATOM 5413 N VAL 76445.741 31.869 53.118 1.00 13.21 0 ATOM 5415 CA VAL 764 45.340 33.14053.680 1.00 13.65 0 ATOM 5416 CB VAL 764 43.825 33.165 53.953 1.00 2.000 ATOM 5417 CG1 VAL 764 43.452 32.097 54.933 1.00 2.00 0 ATOM 5418 CG2VAL 764 43.404 34.521 54.434 1.00 2.00 0 ATOM 5419 C VAL 764 46.15933.363 54.955 1.00 18.12 0 ATOM 5420 O VAL 764 46.396 32.437 55.737 1.002.00 0 ATOM 5421 N THR 765 46.646 34.587 55.111 1.00 2.00 0 ATOM 5423 CATHR 765 47.453 34.992 56.254 1.00 2.00 0 ATOM 5424 CB THR 765 48.73135.660 55.798 1.00 2.00 0 ATOM 5425 OG1 THR 765 49.474 34.740 54.9911.00 2.00 0 ATOM 5427 CG2 THR 765 49.542 36.119 56.973 1.00 2.00 0 ATOM5428 C THR 765 46.675 36.024 57.030 1.00 2.00 0 ATOM 5429 O THR 76546.201 37.016 56.454 1.00 2.00 0 ATOM 5430 N LEU 766 46.549 35.79758.333 1.00 5.65 0 ATOM 5432 CA LEU 766 45.832 36.717 59.205 1.00 5.65 0ATOM 5433 CB LEU 766 44.848 35.943 60.059 1.00 2.00 0 ATOM 5434 CG LEU766 43.964 34.922 59.372 1.00 2.00 0 ATOM 5435 CD1 LEU 766 43.703 33.79460.327 1.00 2.00 0 ATOM 5436 CD2 LEU 766 42.672 35.581 58.943 1.00 2.000 ATOM 5437 C LEU 766 46.826 37.382 60.142 1.00 5.65 0 ATOM 5438 O LEU766 47.864 36.790 60.478 1.00 2.00 0 ATOM 5439 N PHE 767 46.520 38.60560.554 1.00 2.00 0 ATOM 5441 CA PHE 767 47.342 39.320 61.530 1.00 2.00 0ATOM 5442 CB PHE 767 48.259 40.328 60.864 1.00 2.00 0 ATOM 5443 CG PHE767 49.494 40.612 61.649 1.00 2.00 0 ATOM 5444 CD1 PHE 767 50.523 39.67861.695 1.00 2.00 0 ATOM 5445 CD2 PHE 767 49.647 41.811 62.317 1.00 2.000 ATOM 5446 CE1 PHE 767 51.689 39.935 62.389 1.00 2.00 0 ATOM 5447 CE2PHE 767 50.813 42.078 63.018 1.00 2.00 0 ATOM 5448 CZ PHE 767 51.83841.134 63.050 1.00 2.00 0 ATOM 5449 C PHE 767 46.325 40.033 62.418 1.002.00 0 ATOM 5450 O PHE 767 45.957 41.184 62.171 1.00 2.00 0 ATOM 5451 NSER 768 45.850 39.317 63.432 1.00 2.00 0 ATOM 5453 CA SER 768 44.83339.819 64.334 1.00 2.00 0 ATOM 5454 CB SER 768 44.247 38.645 65.091 1.002.00 0 ATOM 5455 OG SER 768 43.903 37.635 64.175 1.00 2.00 0 ATOM 5457 CSER 768 45.261 40.872 65.336 1.00 2.00 0 ATOM 5458 O SER 768 46.29740.731 65.973 1.00 2.00 0 ATOM 5459 N ALA 769 44.435 41.911 65.474 1.0030.20 0 ATOM 5461 CA ALA 769 44.619 43.009 66.428 1.00 27.69 0 ATOM 5462CB ALA 769 45.142 42.454 67.786 1.00 4.86 0 ATOM 5463 C ALA 769 45.41044.253 66.018 1.00 31.35 0 ATOM 5464 O ALA 769 44.855 45.347 65.940 1.0011.84 0 ATOM 5465 N PRO 770 46.717 44.109 65.786 1.00 14.07 0 ATOM 5466CD PRO 770 47.556 42.904 65.939 1.00 2.00 0 ATOM 5467 CA PRO 770 47.57845.209 65.398 1.00 14.07 0 ATOM 5468 CB PRO 770 48.615 44.494 64.5401.00 2.00 0 ATOM 5469 CG PRO 770 48.913 43.329 65.392 1.00 2.00 0 ATOM5470 C PRO 770 47.227 46.584 64.825 1.00 14.07 0 ATOM 5471 O PRO 77046.081 47.016 64.640 1.00 2.00 0 ATOM 5472 N ASN 771 48.363 47.25964.718 1.00 2.00 0 ATOM 5474 CA ASN 771 48.703 48.578 64.220 1.00 2.00 0ATOM 5475 CB ASN 771 48.014 49.664 65.022 1.00 40.66 0 ATOM 5476 CG ASN771 48.270 51.032 64.457 1.00 47.17 0 ATOM 5477 OD1 ASN 771 49.38251.557 64.550 1.00 40.36 0 ATOM 5478 ND2 ASN 771 47.249 51.617 63.8411.00 43.79 0 ATOM 5481 C ASN 771 50.174 48.414 64.666 1.00 2.00 0 ATOM5482 O ASN 771 50.811 49.318 65.208 1.00 50.90 0 ATOM 5483 N TYR 77250.657 47.191 64.417 1.00 6.11 0 ATOM 5485 CA TYR 772 51.955 46.64564.768 1.00 6.11 0 ATOM 5486 CB TYR 772 52.366 45.641 63.705 1.00 8.06 0ATOM 5487 CG TYR 772 53.253 44.546 64.228 1.00 8.06 0 ATOM 5488 CD1 TYR772 52.992 43.949 65.453 1.00 8.06 0 ATOM 5489 CE1 TYR 772 53.781 42.91065.929 1.00 8.06 0 ATOM 5490 CD2 TYR 772 54.335 44.084 63.487 1.00 8.060 ATOM 5491 CE2 TYR 772 55.135 43.048 63.945 1.00 8.06 0 ATOM 5492 CZTYR 772 54.854 42.459 65.170 1.00 8.06 0 ATOM 5493 OH TYR 772 55.63441.403 65.632 1.00 8.06 0 ATOM 5495 C TYR 772 53.130 47.557 65.074 1.006.11 0 ATOM 5496 O TYR 772 53.499 48.428 64.271 1.00 19.22 0 ATOM 5497 NCYS 773 53.724 47.335 66.244 1.00 15.64 0 ATOM 5499 CA CYS 773 54.86848.107 66.681 1.00 13.08 0 ATOM 5500 CB CYS 773 56.059 47.798 65.7771.00 21.47 0 ATOM 5501 SG CYS 773 56.646 46.112 65.891 1.00 23.28 0 ATOM5502 C CYS 773 54.624 49.617 66.685 1.00 16.93 0 ATOM 5503 O CYS 77355.573 50.398 66.832 1.00 20.82 0 ATOM 5504 N GLY 774 53.362 50.03166.545 1.00 2.00 0 ATOM 5506 CA GLY 774 53.059 51.452 66.482 1.00 2.00 0ATOM 5507 C GLY 774 53.821 52.023 65.288 1.00 2.00 0 ATOM 5508 O GLY 77454.051 53.233 65.188 1.00 60.37 0 ATOM 5509 N GLU 775 54.219 51.13064.383 1.00 22.26 0 ATOM 5511 CA GLU 775 54.973 51.500 63.211 1.00 18.010 ATOM 5512 CB GLU 775 56.253 50.683 63.133 1.00 40.50 0 ATOM 5513 CGGLU 775 57.103 50.811 64.357 1.00 44.10 0 ATOM 5514 CD GLU 775 58.49650.246 64.190 1.00 42.58 0 ATOM 5515 OE1 GLU 775 59.404 50.734 64.8941.00 46.10 0 ATOM 5516 OE2 GLU 775 58.687 49.324 63.367 1.00 44.55 0ATOM 5517 C GLU 775 54.181 51.268 61.945 1.00 16.06 0 ATOM 5518 O GLU775 54.133 52.117 61.051 1.00 34.47 0 ATOM 5519 N PHE 776 53.530 50.12561.874 1.00 21.64 0 ATOM 5521 CA PHE 776 52.809 49.796 60.667 1.00 19.450 ATOM 5522 CB PHE 776 52.828 48.279 60.517 1.00 7.64 0 ATOM 5523 CG PHE776 54.213 47.739 60.363 1.00 6.06 0 ATOM 5524 CD1 PHE 776 54.056 47.65661.452 1.00 6.54 0 ATOM 5525 CD2 PHE 776 54.692 47.363 59.122 1.00 10.090 ATOM 5526 CE1 PHE 776 56.370 47.206 61.313 1.00 6.06 0 ATOM 5527 CE2PHE 776 56.005 46.910 58.969 1.00 9.96 0 ATOM 5528 CZ PHE 776 56.84546.833 60.071 1.00 6.06 0 ATOM 5529 C PHE 776 51.424 50.402 60.480 1.0021.71 0 ATOM 5530 O PHE 776 50.979 50.601 59.346 1.00 9.78 0 ATOM 5531 NASP 777 50.757 50.720 61.584 1.00 47.34 0 ATOM 5533 CA ASP 777 49.42751.322 61.532 1.00 47.73 0 ATOM 5534 CB ASP 777 49.526 52.790 61.0711.00 36.82 0 ATOM 5535 CG ASP 777 50.299 53.677 62.059 1.00 86.03 0 ATOM5536 OD1 ASP 777 51.519 53.464 62.246 1.00 85.84 0 ATOM 5537 OD2 ASP 77749.685 54.595 62.644 1.00 85.92 0 ATOM 5538 C ASP 777 48.456 50.54760.632 1.00 47.70 0 ATOM 5539 O ASP 777 47.458 51.095 60.164 1.00 37.690 ATOM 5540 N ASN 778 48.756 49.268 60.419 1.00 12.97 0 ATOM 5542 CA ASN778 47.948 48.373 59.584 1.00 3.97 0 ATOM 5543 CB ASN 778 48.760 47.12359.221 1.00 17.34 0 ATOM 5544 CG ASN 778 49.081 46.258 60.435 1.00 11.190 ATOM 5545 OD1 ASN 778 49.928 46.613 61.275 1.00 10.18 0 ATOM 5546 ND2ASN 778 48.404 45.120 60.538 1.00 7.69 0 ATOM 5549 C ASN 778 46.67247.945 60.296 1.00 4.32 0 ATOM 5550 O ASN 778 46.497 48.227 61.481 1.0021.92 0 ATOM 5551 N ALA 779 45.783 47.267 59.578 1.00 6.26 0 ATOM 5553CA ALA 779 44.537 46.799 60.168 1.00 6.26 0 ATOM 5554 CB ALA 779 43.40446.923 59.174 1.00 78.54 0 ATOM 5555 C ALA 779 44.727 45.344 60.580 1.006.26 0 ATOM 5556 O ALA 779 45.537 44.626 59.988 1.00 86.19 0 ATOM 5557 NGLY 780 43.998 44.925 61.606 1.00 2.00 0 ATOM 5559 CA GLY 780 44.09143.560 62.077 1.00 2.00 0 ATOM 5560 C GLY 780 42.800 42.848 61.747 1.002.00 0 ATOM 5561 O GLY 780 41.725 43.433 61.869 1.00 21.77 0 ATOM 5562 NALA 781 42.880 41.587 61.345 1.00 2.00 0 ATOM 5564 CA ALA 781 41.67040.870 60.984 1.00 2.00 0 ATOM 5565 CB ALA 781 41.671 40.567 59.492 1.0014.58 0 ATOM 5566 C ALA 781 41.474 39.587 61.763 1.00 2.00 0 ATOM 5567 OALA 781 42.396 39.076 62.398 1.00 14.58 0 ATOM 5568 N MET 782 40.26339.061 61.660 1.00 2.00 0 ATOM 5570 CA MET 782 39.857 37.833 62.319 1.002.00 0 ATOM 5571 CB MET 782 39.037 38.204 63.554 1.00 2.00 0 ATOM 5572CG MET 782 38.736 37.067 64.502 1.00 2.00 0 ATOM 5573 SD MET 782 37.74337.608 65.912 1.00 2.00 0 ATOM 5574 CE MET 782 37.595 39.370 65.621 1.002.00 0 ATOM 5575 C MET 782 39.010 37.029 61.306 1.00 2.00 0 ATOM 5576 OMET 782 38.090 37.570 60.688 1.00 2.00 0 ATOM 5577 N MET 783 39.32535.752 61.116 1.00 2.00 0 ATOM 5579 CA MET 783 38.570 34.955 60.165 1.002.00 0 ATOM 5580 CB MET 783 39.482 34.182 59.209 1.00 16.43 0 ATOM 5581CG MET 783 38.688 33.363 58.165 1.00 19.34 0 ATOM 5582 SD MET 783 39.68932.337 57.060 1.00 18.69 0 ATOM 5583 CE MET 783 39.780 30.793 57.9751.00 21.53 0 ATOM 5584 C MET 783 37.608 33.976 60.794 1.00 2.00 0 ATOM5585 O MET 783 38.004 33.010 61.447 1.00 10.33 0 ATOM 5586 N SER 78436.335 34.221 60.542 1.00 2.00 0 ATOM 5588 CA SER 784 35.276 33.37461.034 1.00 2.00 0 ATOM 5589 CB SER 784 34.033 34.220 61.308 1.00 16.250 ATOM 5590 OG SER 784 34.385 35.392 62.020 1.00 16.25 0 ATOM 5592 C SER784 34.953 32.301 59.990 1.00 2.00 0 ATOM 5593 O SER 784 34.672 32.61158.839 1.00 19.28 0 ATOM 5594 N VAL 785 35.033 31.045 60.401 1.00 10.880 ATOM 5596 CA VAL 785 34.706 29.907 59.557 1.00 16.53 0 ATOM 5597 CBVAL 785 35.649 28.739 59.792 1.00 11.43 0 ATOM 5598 CG1 VAL 785 35.18327.538 58.975 1.00 11.43 0 ATOM 5599 CG2 VAL 785 37.082 29.153 59.4771.00 11.43 0 ATOM 5600 C VAL 785 33.357 29.465 60.083 1.00 12.38 0 ATOM5601 O VAL 785 33.254 28.998 61.225 1.00 11.43 0 ATOM 5602 N ASP 78632.307 29.613 59.291 1.00 2.00 0 ATOM 5604 CA ASP 786 31.024 29.20059.807 1.00 2.00 0 ATOM 5605 CB ASP 786 29.874 30.008 59.171 1.00 24.900 ATOM 5606 CG ASP 786 28.403 29.458 57.842 1.00 30.71 0 ATOM 5607 OD1ASP 786 28.245 29.765 57.474 1.00 33.80 0 ATOM 5608 OD2 ASP 786 30.16528.737 57.163 1.00 32.56 0 ATOM 5609 C ASP 786 30.860 27.699 59.656 1.002.00 0 ATOM 5610 O ASP 786 31.677 27.029 59.031 1.00 23.30 0 ATOM 5611 NGLU 787 29.803 27.195 60.268 1.00 7.48 0 ATOM 5613 CA GLU 787 29.44725.782 60.270 1.00 13.41 0 ATOM 5614 CB GLU 787 27.983 25.636 60.6961.00 2.00 0 ATOM 5615 CG GLU 787 27.174 26.963 60.747 1.00 2.00 0 ATOM5616 CD GLU 787 27.384 27.758 62.053 1.00 2.00 0 ATOM 5617 OE1 GLU 78726.858 27.319 63.117 1.00 2.00 0 ATOM 5618 OE2 GLU 787 28.070 28.81762.024 1.00 2.00 0 ATOM 5619 C GLU 787 29.665 25.015 58.965 1.00 11.86 0ATOM 5620 O GLU 787 30.002 23.830 58.994 1.00 2.00 0 ATOM 5621 N THR 78829.492 25.698 57.836 1.00 56.74 0 ATOM 5623 CA THR 788 29.616 25.09256.513 1.00 52.70 0 ATOM 5624 CB THR 788 28.369 25.402 55.700 1.00 6.460 ATOM 5625 OG1 THR 788 28.317 26.819 55.467 1.00 4.65 0 ATOM 5627 CG2THR 788 27.101 24.968 56.457 1.00 10.48 0 ATOM 5628 C THR 788 30.83525.536 55.683 1.00 51.86 0 ATOM 5629 O THR 788 30.751 25.628 54.449 1.0012.46 0 ATOM 5630 N LEU 789 31.946 25.823 56.364 1.00 10.40 0 ATOM 5632CA LEU 789 33.194 26.245 55.734 1.00 6.34 0 ATOM 5633 CB LEU 789 33.67025.165 54.775 1.00 8.80 0 ATOM 5634 CG LEU 789 34.458 24.017 55.405 1.0015.93 0 ATOM 5635 CD1 LEU 789 35.879 24.487 55.677 1.00 11.55 0 ATOM5636 CD2 LEU 789 33.785 23.525 56.682 1.00 14.98 0 ATOM 5637 C LEU 78933.173 27.619 55.042 1.00 4.53 0 ATOM 5638 O LEU 789 34.065 27.94854.248 1.00 8.97 0 ATOM 5639 N MET 790 32.165 28.430 55.336 1.00 2.00 0ATOM 5641 CA MET 790 32.126 29.748 54.743 1.00 2.00 0 ATOM 5642 CB MET790 30.698 30.267 54.636 1.00 19.79 0 ATOM 5643 CG MET 790 30.588 31.57253.882 1.00 19.37 0 ATOM 5644 SD MET 790 28.979 31.708 53.134 1.00 18.510 ATOM 5645 CE MET 790 28.359 33.110 53.992 1.00 23.23 0 ATOM 5646 C MET790 32.945 30.667 55.627 1.00 2.00 0 ATOM 5647 O MET 790 32.615 30.85756.799 1.00 23.79 0 ATOM 5648 N CYS 791 34.014 31.228 55.067 1.00 2.00 0ATOM 5650 CA CYS 791 34.882 32.133 55.803 1.00 2.00 0 ATOM 5651 CB CYS791 36.325 31.793 55.516 1.00 14.83 0 ATOM 5652 SG CYS 791 36.570 30.05255.766 1.00 15.43 0 ATOM 5653 C CYS 791 34.616 33.591 55.502 1.00 2.00 0ATOM 5654 O CYS 791 34.314 33.961 54.371 1.00 21.84 0 ATOM 5655 N SER792 34.697 34.412 56.540 1.00 2.00 0 ATOM 5657 CA SER 792 34.480 35.84956.430 1.00 2.00 0 ATOM 5658 CB SER 792 33.073 36.221 56.903 1.00 2.93 0ATOM 5659 OG SER 792 32.887 35.867 58.256 1.00 3.41 0 ATOM 5661 C SER792 35.539 36.518 57.303 1.00 2.00 0 ATOM 5662 O SER 792 36.290 35.82657.994 1.00 2.56 0 ATOM 5663 N PHE 793 35.609 37.842 57.270 1.00 2.00 0ATOM 5665 CA PHE 793 36.617 38.558 58.039 1.00 2.00 0 ATOM 5666 CB PHE793 37.765 39.025 57.129 1.00 38.05 0 ATOM 5667 CG PHE 793 38.531 37.91356.469 1.00 24.74 0 ATOM 5668 CD1 PHE 793 38.055 37.314 55.306 1.0028.08 0 ATOM 5669 CD2 PHE 793 39.741 37.474 57.002 1.00 26.11 0 ATOM5670 CE1 PHE 793 38.774 36.295 54.680 1.00 27.84 0 ATOM 5671 CE2 PHE 79340.466 36.460 56.389 1.00 27.16 0 ATOM 5672 CZ PHE 793 39.983 35.86855.225 1.00 29.41 0 ATOM 5673 C PHE 793 36.101 39.782 58.784 1.00 2.00 0ATOM 5674 O PHE 793 35.362 40.599 58.233 1.00 23.22 0 ATOM 5675 N GLN794 36.480 39.893 60.051 1.00 67.38 0 ATOM 5677 CA GLN 794 36.128 41.06460.837 1.00 63.52 0 ATOM 5678 CB GLN 794 35.608 40.713 62.238 1.00 31.740 ATOM 5679 CG GLN 794 34.294 39.954 62.289 1.00 36.03 0 ATOM 5680 CDGLN 794 34.506 38.455 62.252 1.00 39.99 0 ATOM 5681 OE1 GLN 794 34.62237.858 61.179 1.00 39.14 0 ATOM 5682 NE2 GLN 794 34.575 37.838 63.4251.00 44.72 0 ATOM 5685 C GLN 794 37.471 41.760 60.958 1.00 65.96 0 ATOM5686 O GLN 794 38.486 41.132 61.272 1.00 37.24 0 ATOM 5687 N ILE 79537.498 43.045 60.658 1.00 53.07 0 ATOM 5689 CA ILE 795 38.732 43.79060.748 1.00 54.05 0 ATOM 5690 CB ILE 795 39.084 44.483 59.392 1.00 26.190 ATOM 5691 CG2 ILE 795 40.115 45.589 59.608 1.00 28.70 0 ATOM 5692 CG1ILE 795 39.643 43.467 58.390 1.00 30.47 0 ATOM 5693 CD1 ILE 795 38.65942.449 57.906 1.00 28.83 0 ATOM 5694 C ILE 795 38.543 44.847 61.817 1.0053.94 0 ATOM 5695 O ILE 795 37.476 45.452 61.903 1.00 27.08 0 ATOM 5696N LEU 796 39.544 45.008 62.674 1.00 30.60 0 ATOM 5698 CA LEU 796 39.52146.054 63.686 1.00 34.91 0 ATOM 5699 CB LEU 796 39.839 45.528 65.0891.00 41.88 0 ATOM 5700 CG LEU 796 40.368 44.121 65.308 1.00 41.67 0 ATOM5701 CD1 LEU 796 40.976 44.038 66.702 1.00 39.26 0 ATOM 5702 CD2 LEU 79639.242 43.114 65.119 1.00 42.51 0 ATOM 5703 C LEU 796 40.637 46.97763.206 1.00 33.78 0 ATOM 5704 O LEU 796 41.649 46.510 62.667 1.00 34.590 ATOM 5705 N LYS 797 40.454 48.280 63.356 1.00 37.84 0 ATOM 5707 CA LYS797 41.475 49.207 62.902 1.00 40.12 0 ATOM 5708 CB LYS 797 40.805 50.35462.154 1.00 0.26 0 ATOM 5709 CG LYS 797 39.959 49.929 60.932 1.00 0.34 0ATOM 5710 CD LYS 797 39.456 51.183 60.151 1.00 0.65 0 ATOM 5711 CE LYS797 39.134 50.875 58.662 1.00 0.12 0 ATOM 5712 NZ LYS 797 38.852 52.11757.851 1.00 0.70 0 ATOM 5716 C LYS 797 42.356 49.727 64.053 1.00 50.42 0ATOM 5717 O LYS 797 42.961 50.809 63.985 1.00 0.89 0 ATOM 5718 N ALA 400−8.399 33.628 131.469 1.00 75.56 0 ATOM 5720 CA ALA 400 −6.981 33.565131.818 1.00 75.56 0 ATOM 5721 CB ALA 400 −6.134 33.160 130.576 1.0021.27 0 ATOM 5722 C ALA 400 −6.530 34.922 132.352 1.00 75.56 0 ATOM 5723O ALA 400 −7.350 35.762 132.744 1.00 21.27 0 ATOM 5724 N ARG 401 −5.21835.116 132.374 1.00 2.00 0 ATOM 5726 CA ARG 401 −4.619 36.351 132.8331.00 2.00 0 ATOM 5727 CB ARG 401 −4.586 36.395 134.359 1.00 2.00 0 ATOM5728 CG ARG 401 −5.638 37.360 134.956 1.00 2.00 0 ATOM 5729 CD ARG 401−5.639 37.340 136.471 1.00 2.00 0 ATOM 5730 NE ARG 401 −6.285 38.512137.058 1.00 2.00 0 ATOM 5732 CZ ARG 401 −5.730 39.260 138.022 1.00 2.000 ATOM 5733 NH1 ARG 401 −6.338 40.369 138.457 1.00 2.00 0 ATOM 5736 NH2ARG 401 −4.527 38.949 138.511 1.00 2.00 0 ATOM 5739 C ARG 401 −3.21636.446 132.267 1.00 2.00 0 ATOM 5740 O ARG 401 −2.505 35.443 132.1501.00 2.00 0 ATOM 5741 N VAL 402 −2.822 37.655 131.898 1.00 2.00 0 ATOM5743 CA VAL 402 −1.516 37.882 131.311 1.00 2.00 0 ATOM 5744 CB VAL 402−1.331 39.379 131.042 1.00 2.00 0 ATOM 5745 CG1 VAL 402 −0.063 39.632130.283 1.00 2.00 0 ATOM 5746 CG2 VAL 402 −2.503 39.889 130.282 1.002.00 0 ATOM 5747 C VAL 402 −0.398 37.376 132.223 1.00 2.00 0 ATOM 5748 OVAL 402 −0.548 37.369 133.444 1.00 2.00 0 ATOM 5749 N SER 403 0.70136.920 131.625 1.00 19.26 0 ATOM 5751 CA SER 403 1.882 36.461 132.3611.00 23.92 0 ATOM 5752 CB SER 403 1.894 34.942 132.528 1.00 22.54 0 ATOM5753 OG SER 403 1.503 34.298 131.333 1.00 14.30 0 ATOM 5755 C SER 4033.070 36.913 131.514 1.00 21.44 0 ATOM 5756 O SER 403 2.890 37.720130.595 1.00 23.72 0 ATOM 5757 N PHE 404 4.271 36.415 131.793 1.00 13.470 ATOM 5759 CA PHE 404 5.439 36.827 131.013 1.00 13.06 0 ATOM 5760 CBPHE 404 6.101 38.060 131.647 1.00 2.00 0 ATOM 5761 CG PHE 404 5.22839.270 131.627 1.00 2.00 0 ATOM 5762 CD1 PHE 404 4.421 39.574 132.7111.00 2.00 0 ATOM 5763 CD2 PHE 404 5.171 40.080 130.505 1.00 2.00 0 ATOM5764 CE1 PHE 404 3.560 40.668 132.677 1.00 2.00 0 ATOM 5765 CE2 PHE 4044.313 41.178 130.463 1.00 2.00 0 ATOM 5766 CZ PHE 404 3.507 41.472131.549 1.00 2.00 0 ATOM 5767 C PHE 404 6.476 35.735 130.791 1.00 16.540 ATOM 5768 O PHE 404 6.472 34.720 131.489 1.00 2.00 0 ATOM 5769 N ALA405 7.341 35.960 129.802 1.00 2.00 0 ATOM 5771 CA ALA 405 8.420 35.054129.414 1.00 2.00 0 ATOM 5772 CB ALA 405 9.762 30.742 129.619 1.00 85.030 ATOM 5773 C ALA 405 8.434 33.682 130.078 1.00 2.00 0 ATOM 5774 O ALA405 8.380 32.654 129.401 1.00 85.03 0 ATOM 5775 N GLY 899 32.968 17.22649.661 1.00 95.94 0 ATOM 5777 CA GLY 899 31.781 16.989 50.464 1.00 95.940 ATOM 5778 C GLY 899 31.215 18.274 51.038 1.00 95.94 0 ATOM 5779 O GLY899 30.022 18.368 51.338 1.00 32.75 0 ATOM 5780 N ARG 900 32.090 19.26051.193 1.00 42.23 0 ATOM 5782 CA ARG 900 31.732 20.569 51.722 1.00 42.230 ATOM 5783 CB ARG 900 31.110 20.432 53.110 1.00 22.01 0 ATOM 5784 CGARG 900 30.578 21.718 53.694 1.00 22.01 0 ATOM 5785 CD ARG 900 30.08021.488 55.120 1.00 22.01 0 ATOM 5786 NE ARG 900 31.005 20.664 55.9091.00 22.01 0 ATOM 5788 CZ ARG 900 31.111 20.691 57.237 1.00 22.01 0 ATOM5789 NH1 ARG 900 30.361 21.512 57.963 1.00 22.01 0 ATOM 5792 NH2 ARG 90031.957 19.865 57.839 1.00 22.01 0 ATOM 5795 C ARG 900 33.034 21.36551.774 1.00 42.23 0 ATOM 5796 O ARG 900 33.795 21.309 52.735 1.00 22.010 ATOM 5797 N ARG 901 33.284 22.091 50.697 1.00 8.39 0 ATOM 5799 CA ARG901 34.490 22.880 50.542 1.00 8.39 0 ATOM 5800 CB ARG 901 34.793 23.03649.045 1.00 2.00 0 ATOM 5801 CG ARG 901 34.504 21.810 48.186 1.00 2.00 0ATOM 5802 CD ARG 901 34.584 22.175 46.699 1.00 2.00 0 ATOM 5803 NE ARG901 34.278 21.012 45.863 1.00 2.00 0 ATOM 5805 CZ ARG 901 35.190 20.19545.327 1.00 2.00 0 ATOM 5606 NH1 ARG 901 36.498 20.416 45.509 1.00 2.000 ATOM 5809 NH2 ARG 901 34.791 19.114 44.649 1.00 2.00 0 ATOM 5812 C ARG901 34.327 24.259 51.158 1.00 8.39 0 ATOM 5813 O ARG 901 33.241 24.62951.607 1.00 2.00 0 ATOM 5814 N VAL 902 35.393 25.005 51.149 1.00 2.00 0ATOM 5816 CA VAL 902 35.425 26.384 51.650 1.00 2.00 0 ATOM 5817 CB VAL902 36.880 26.814 51.869 1.00 12.90 0 ATOM 5818 CG1 VAL 902 37.03928.332 51.992 1.00 12.90 0 ATOM 5819 CG2 VAL 902 37.492 26.219 53.1391.00 12.90 0 ATOM 5820 C VAL 902 34.782 27.324 50.627 1.00 2.00 0 ATOM5621 O VAL 902 34.737 27.031 49.435 1.00 12.90 0 ATOM 5822 N SER 90334.288 28.438 51.120 1.00 2.00 0 ATOM 5824 CA SER 903 33.673 29.48750.271 1.00 2.00 0 ATOM 5825 CB SER 903 32.173 29.205 50.008 1.00 2.00 0ATOM 5826 OG SER 903 31.477 28.917 51.209 1.00 2.00 0 ATOM 5828 C SER903 33.871 30.814 50.991 1.00 2.00 0 ATOM 5829 O SER 903 34.386 30.86652.115 1.00 2.00 0 ATOM 5830 N PHE 904 33.482 31.903 50.376 1.00 2.00 0ATOM 5832 CA PHE 904 33.683 33.188 51.034 1.00 2.00 0 ATOM 5833 CB PHE904 34.894 33.898 50.455 1.00 17.14 0 ATOM 5834 CG PHE 904 36.184 33.10950.672 1.00 17.14 0 ATOM 5835 CD1 PHE 904 36.511 32.067 49.605 1.0017.14 0 ATOM 5836 CD2 PHE 904 37.034 33.426 51.738 1.00 17.14 0 ATOM5837 CE1 PHE 904 37.690 31.343 49.994 1.00 17.14 0 ATOM 5838 CE2 PHE 90438.216 32.704 51.927 1.00 17.14 0 ATOM 5839 CZ PHE 904 38.544 31.66251.054 1.00 17.14 0 ATOM 5840 C PHE 904 32.491 34.080 50.894 1.00 2.00 0ATOM 5841 O PHE 904 31.716 33.994 49.926 1.00 17.14 0 ATOM 5842 N ALA905 32.386 34.912 51.874 1.00 70.83 0 ATOM 5844 CA ALA 905 31.311 35.85751.949 1.00 69.99 0 ATOM 5845 CB ALA 905 30.042 35.128 52.370 1.00 2.000 ATOM 5846 C ALA 905 31.646 36.940 52.962 1.00 69.85 0 ATOM 5847 O ALA905 30.981 37.068 53.993 1.00 2.00 0 ATOM 5848 N ALA 907 37.374 34.31247.285 1.00 23.39 0 ATOM 5850 CA ALA 907 36.215 33.651 46.695 1.00 23.390 ATOM 5851 CB ALA 907 35.317 34.681 46.008 1.00 41.64 0 ATOM 5852 C ALA907 36.637 32.568 45.699 1.00 23.39 0 ATOM 5853 O ALA 907 35.850 31.68045.370 1.00 41.64 0 ATOM 5854 N ALA 908 37.875 32.643 45.215 1.00 83.900 ATOM 5856 CA ALA 908 38.378 31.663 44.253 1.00 83.90 0 ATOM 5857 CBALA 908 39.439 32.296 43.337 1.00 22.84 0 ATOM 5858 C ALA 908 38.96130.466 44.992 1.00 83.90 0 ATOM 5859 O ALA 908 38.459 30.074 46.046 1.0022.84 0 ATOM 5860 OW WAT 1 62.869 37.982 63.341 1.00 20.00 0 ATOM 5863OW WAT 103 57.039 39.062 61.228 1.00 20.00 0 ATOM 5866 OW WAT 101 7.25766.194 118.365 1.00 20.00 0 ATOM 5869 OW WAT 2 12.341 60.209 123.4641.00 20.00 0 ATOM 5872 OW WAT 1 10.655 60.748 120.831 1.00 20.00 0 ATOM5875 OW WAT 104 55.432 36.306 63.901 1.00 20.00 0 ATOM 5878 OW WAT 10256.782 40.457 58.333 1.00 20.00 0 ATOM 5881 OW WAT 4 5.787 57.856118.686 1.00 20.00 0 ATOM 5884 OW WAT 105 54.382 39.155 63.734 1.0020.00 0 ATOM 5887 OW WAT 5 8.964 57.595 118.151 1.00 20.00 0 ATOM 5890OW WAT 106 38.565 47.423 74.959 1.00 20.00 0 ATOM 5893 OW WAT 6 16.08642.169 105.289 1.00 20.00 0 ATOM 5896 OW WAT 107 31.158 26.414 51.9131.00 20.00 0 ATOM 5899 OW WAT 7 −0.781 32.787 131.574 1.00 20.00 0 ATOM5902 MN MN2 430 4.422 59.061 119.360 1.00 15.61 0 ATOM 5903 MN MN2 4317.458 57.871 117.661 1.00 16.53 0 ATOM 5904 MN MN2 930 56.038 34.50063.727 1.00 16.67 0 ATOM 5905 MN MN2 931 54.402 37.798 64.756 1.00 15.400 ATOM 5906 S SO4 801 57.551 37.278 64.009 1.00 37.87 0 ATOM 5907 O1 SO4801 57.600 35.652 63.897 1.00 42.46 0 ATOM 5908 O2 SO4 801 58.690 37.74064.722 1.00 42.01 0 ATOM 5909 O3 SO4 801 56.355 37.648 64.705 1.00 45.080 ATOM 5910 O4 SO4 801 57.520 37.854 62.725 1.00 41.40 0 ATOM 5911 S SO4800 6.866 60.776 118.643 1.00 37.87 0 ATOM 5912 O1 SO4 800 7.710 60.635119.773 1.00 42.46 0 ATOM 5913 O2 SO4 800 7.044 62.063 118.053 1.0042.01 0 ATOM 5914 O3 SO4 800 5.496 60.612 119.046 1.00 45.08 0 ATOM 5915O4 SO4 800 7.194 59.728 117.703 1.00 41.40 0

References for Example 3

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References (Given as Numbers in Square Brackets, Except in Example 3)

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1. A method of identifying a compound which modulates the interactionbetween a PP1c and a regulatory subunit thereof, the method comprisingassaying for the interaction between (a) a PP1c and (b) a regulatorysubunit which is able to bind to PP1c or a PP1c-binding fragment orvariant, wherein the fragment or variant comprises SEQ ID NO:35, orfusion of said subunit or a fusion of said fragment or variant, whereinthe regulatory subunit is any one of M₁₁₀, G_(L), G_(M), M-complexes,p53 BP2, sds22, NIPPI, L5, Inhibitor-1, Inhibitor-2, or DARPP, andassaying for the interaction in the presence of a compound, to identifya compound which modulates the interaction.
 2. A method of identifying acompound which mimics the effect of a regulatory subunit of PP1c, themethod comprising assaying for the enzymatic or binding activity of PP1cin the presence of a given regulatory subunit, and contacting a compoundwith PP1c and determining whether, in the presence of the compound, PP1cadopts the enzymatic or binding activity of the PP1c in the presence ofthe given regulatory subunit, wherein if PP1c adopts the enzymatic orbinding activity of the PP1c in the presence of a given regulatorysubunit, the compound mimics the effect of the regulatory subunit ofPP1c, and wherein the regulatory subunit is any one of M₁₁₀, G_(L),G_(M), M-complexes, p53 BP2, sds22, NIPPI, L5, Inhibitor-1, Inhibitor-2,or DARPP.
 3. A method according to claim 1 or 2 wherein the regulatorysubunit of PP1c is any one of M₁₁₀, G_(L), G_(M), M-complexes or p53BP2.4. A method according to claim 3 wherein the regulatory subunit of PP1cis M₁₁₀ or G_(M).
 5. A method according to claim 1 wherein the fragmentof a regulatory subunit which is able to bind to PP1c is any of thepeptides G63-T93 of SEQ ID NO:32, G63-N75 of SEQ ID NO:32, E2-P243 ofSEQ ID NO:34, E2-D118 of SEQ ID NO:34, and peptide 63-80 of SEQ ID NO:32G_(M) or peptides comprising said peptide sequences provided that theyare not the complete G_(M) regulatory subunit.
 6. A method according toclaim 1 wherein the fragment of a regulatory subunit which is able tobind to PP1c is any of the peptides M1-E309 of SEQ ID NO:33, M1-F38 ofSEQ ID NO:33, M1-A150 of SEQ ID NO:33, or L24-Y496 of SEQ ID NO:33 ofM₁₁₀ or peptides comprising said peptide sequences provided that theyare not the complete M₁₁₀ regulatory subunit.
 7. A method according toclaim 1 wherein the PP1c-binding fragment or variant further comprisesat least one basic residue in the four residues N-terminal of theconsensus peptide sequence.
 8. A method according to claim 1, whereinthe third amimo acid in SEQ ID NO:35 is not Asp or Glu or a largehydrophobic residue.
 9. A method according to claim 1 wherein thePP1c-binding fragment is a fragment of a regulatory subunit.
 10. Amethod according to claim 8 wherein the PP1c-binding fragment is afragment of any of the M₁₁₀, G_(L), G_(M), M-complexes, p53BP2, sds22,NIPPI, L5, Inhibitor-1, Inhibitor-2 or DARPP regulatory subunitscomprising said SEQ ID NO:35.
 11. A method according to claim 1 or 2wherein the compound binds to a PP1c.
 12. A method according to claim 1wherein the compound binds to a regulatory subunit of PP1c.
 13. Acompound which modulates the interaction between a PP1c and a regulatorysubunit thereof said compound comprising any of the peptides G63-T93 ofSEQ ID NO:32, G63-N75 of SEQ ID NO:32, E2-P243 of SEQ ID NO:34, E2-D118of SEQ ID NO:34, and peptide 63-80 of SEQ ID NO:32 G_(M) or saidcompound comprising any of the peptides M1-E309 of SEQ ID NO:33, M1-F38of SEQ ID NO:33, M1-A150 of SEQ ID NO:33 or L24-Y496 of SEQ ID NO:33 ofM₁₁₀ or said compound comprising the consensus peptide sequence SEQ IDNO:35:Arg/Lys-Val/Ile-Xaa-Phe wherein Xaa is any naturally occurringamino acid, provided that said compound is not a complete regulatorysubunit of PP1c.
 14. A compound according to claim 13 consisting of thepeptides G63-T93 of SEQ ID NO:32, G63-N75 of SEQ ID NO:32, E2-P243 ofSEQ ID NO:34, E2-D118 of SEQ ID NO:34, or peptide 63-80 of SEQ ID NO:32G_(M) or consisting of the peptides M1-E309 of SEQ ID NO:33, M1-F38 ofSEQ ID NO:33, M1-A150 of SEQ ID NO:33 or L24-Y496 of SEQ ID NO:33 ofM₁₁₀.
 15. A method of identifying a compound which modulates theinteraction between a PP1c and a regulatory subunit thereof, or bindsPP1c or mimics the effect of a regulatory subunit, the method comprisingdetermining the conformation of a peptide bound to a regulatorysubunit-binding site of PP1c and the conformation of the portion of PP1cwhich binds to the peptide, and selecting a compound which is capable ofadopting the same conformation as the peptide bound to the regulatorysubunit-binding site of PP1c or the same conformation as the portion ofPP1c which binds to said peptide, wherein the selected compoundmodulates the interaction between a PP1c and a regulatory subunitthereof or binds PP1c and a regulatory subunit thereof or binds PP1c ormimics the effect of a regulatory subunit, wherein the regulatorysubunit is any one of M₁₁₀, G_(L), G_(M), M-complexes, p53BP2, sds22,NIPPI, L5, Inhibitor-1, Inhibitor-2 or DARPP.
 16. A method according toclaim 15 wherein said peptide comprises the consensus peptide sequenceSEQ ID NO:35: Arg/Lys-Val/Ile-Xaa-Phe wherein Xaa is any amino acid. 17.A method according to claim 16 wherein said peptide consists of residues63 to 75 of SEQ ID NO: 32 G_(m).
 18. A composition comprising a compoundaccording to claim 13 and an acceptable carrier.
 19. A method accordingto claim 2 wherein the function or properties of the PP1c include thesubcellular location of the PP1c, the substrate specificity of the PP1c,the activity of the PP1c towards one or more substrates, the activity ofthe PP1c in the presence of one or more extracellular agonists, theability of the PP1c to bind to a regulatory subunit, or the ability ofthe PP1c to be modulated by reversible protein phosphorylation or secondmessengers.